N1-PYRAZOLOSPIROKETONE ACETYL-CoA CARBOXYLASE INHIBITORS

ABSTRACT

The invention provides a compound of Formula (I) 
     
       
         
         
             
             
         
       
     
     or a pharmaceutically acceptable salt of the compound, wherein R 1 , R 2 , R 3 , Z, A 1 , L and A 2  are as described herein; pharmaceutical compositions thereof; and the use thereof in treating diseases, conditions or disorders modulated by the inhibition of an acetyl-CoA carboxylase enzyme(s) in an animal.

FIELD OF THE INVENTION

This invention relates to substituted pyrazolospiroketone compounds thatact as inhibitors of an acetyl-CoA carboxylase(s) and their use intreating diseases, conditions or disorders modulated by the inhibitionof acetyl-CoA carboxylase enzyme(s).

BACKGROUND OF THE INVENTION

Acetyl-CoA carboxylases (ACC) are a family of enzymes found in mostspecies and are associated with fatty acid synthesis and metabolismthrough catalyzing the production of malonyl-CoA from acetyl-CoA. Inmammals, two isoforms of the ACC enzyme have been identified. ACC1,which is expressed at high levels in lipogenic tissues, such as fat andthe liver, controls the first committed step in the biosynthesis oflong-chain fatty acids. If acetyl-CoA is not carboxylated to formmalonyl-CoA, it is metabolized through the Krebs cycle. ACC2, a minorcomponent of hepatic ACC but the predominant isoform in heart andskeletal muscle, catalyzes the production of malonyl-CoA at thecytosolic surface of mitochondria, and regulates how much fatty acid isutilized in β-oxidation by inhibiting carnitine palmitoyl transferase.Thus, by increasing fatty acid utilization and by preventing increasesin de novo fatty acid synthesis, chronic administration of an ACCinhibitor (ACC-I) may also deplete liver and adipose tissue triglyceride(TG) stores in obese subjects consuming a high or low-fat diet, leadingto selective loss of body fat.

Studies conducted by Abu-Etheiga, et al., suggest that ACC2 plays anessential role in controlling fatty acid oxidation and, as such it wouldprovide a target in therapy against obesity and obesity-relateddiseases, such as type-2 diabetes. See, Abu-Etheiga, L., et al.,“Acetyl-CoA carboxylase 2 mutant mice are protected against obesity anddiabetes induced by high-fat/high-carbohydrate diets” PNAS, 100(18)10207-10212 (2003). See also, Choi, C. S., et al., “Continuous fatoxidation in acetyl-CoA carboxylase 2 knockout mice increases totalenergy expenditure, reduces fat mass, and improves insulin sensitivity”PNAS, 104(42) 16480-16485 (2007).

It is becoming increasingly clear that hepatic lipid accumulation causeshepatic insulin resistance and contributes to the pathogenesis of type 2diabetes. Salvage, et al., demonstrated that ACC 1 and ACC2 are bothinvolved in regulating fat oxidation in hepatocytes while ACC1, thedominant isoform in rat liver, is the sole regulator of fatty acidsynthesis. Furthermore, in their model, combined reduction of bothisoforms is required to significantly lower hepatic malonyl-CoA levels,increase fat oxidation in the fed state, reduce lipid accumulation, andimprove insulin action in vivo. Thus, showing that hepatic ACC1 and ACC2inhibitors may be useful in the treatment of nonalcoholic fatty liverdisease (NAFLD) and hepatic insulin resistance. See, Savage, D. B., etal., “Reversal of diet-induced hepatic steatosis and hepatic insulinresistance by antisense oligonucleotide inhibitors of acetyl-CoAcarboxylases 1 and 2” J Clin Invest doi: 10.1172/JCI27300. See also, Oh,W., et al., “Glucose and fat metabolism in adipose tissue of acetyl-CoAcarboxylase 2 knockout mice” PNAS, 102(5) 1384-1389 (2005).

Consequently, there is a need for medicaments containing ACC1 and/orACC2 inhibitors to treat obesity and obesity-related diseases (such as,NAFLD and type-2 diabetes) by inhibiting fatty acid synthesis and byincreasing fatty acid oxidation.

SUMMARY OF THE INVENTION

A first embodiment of the present invention is a compound having thestructure of Formula (I)

or a pharmaceutically acceptable salt thereof; wherein

R¹ is (C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, tetrahydrofuranyl or oxetanyl;wherein said (C₁-C₆)alkyl is optionally substituted with 1 to 3substituents independently selected from (C₁-C₃)alkoxy, hydroxy, fluoro,phenyl, tetrahydrofuranyl or oxetanyl;

R² is hydrogen, halo, (C₁-C₃)alkyl, or cyano;

R³ are each independently hydrogen or (C₁-C₃)alkyl;

L is a direct bond or a (C₁-C₆)alkylene wherein one carbon of the(C₁-C₆)alkylene is optionally replaced by —C(O)—, —C(O)NH—, —NHC(O), O,S, NH or N(C₁-C₃)alkyl;

Z is CH₂ or O;

A¹ and A² are each independently (C₆-C₁₀)aryl, 5 to 12 memberedheteroaryl or 8 to 12 membered fused heterocyclicaryl; wherein said(C₆-C₁₀)aryl, 5 to 12 membered heteroaryl or 8 to 12 membered fusedheterocyclicaryl are each optionally substituted with one to threesubstituents independently selected from (C₁-C₃)alkyl, (C₁-C₃)alkoxy,halo, amino, (C₁-C₃)alkylamino, di(C₁-C₃)alkylamino, hydroxy, cyano andamido wherein the alkyl portion of the (C₁-C₃)alkyl, (C₁-C₃)alkoxy,(C₁-C₃)alkylamino and di(C₁-C₃)alkylamino are optionally substitutedwith one to five fluoro; and wherein one of A¹ or A² is substituted byCO₂R⁴, (C₁-C₆)CO₂R⁴, tetrazolyl or (C₁-C₆)tetrazolyl; and

R⁴ is (C₁-C₈)alkyl, (C₃-C₈)cycloalkyl or (C₁-C₆)alkyl-(C₃-C₈)cycloalkyl;or a pharmaceutically acceptable salt thereof.

Another embodiment of the present invention is the compound of theimmediately preceding embodiment wherein R¹ is (C₁-C₆)alkyl,(C₃-C₇)cycloalkyl, or tetrahydrofuranyl; R² is hydrogen or methyl; eachR³ is hydrogen; and L is a direct bond or O; or a pharmaceuticallyacceptable salt thereof.

Another embodiment of the present invention is the compound of theimmediately preceding embodiment wherein R¹ is (C₂-C₄)alkyl; A¹ and A²are each independently phenyl, pyrazolyl, imidazolyl, triazolyl,pyridinyl, pyrimidinyl, indolyl, benzopyrazinyl, benzoimidazolyl,benzoimidazolonyl, pyrrolopyridinyl, pyrrolopyrimidinyl,pyrazolopyridinyl, pyrazolopyrimidinyl, indazolyl, indolinonyl,naphthyridinyl, quinolinyl, quinolinonyl, dihydroquinolinonyl,oxo-dihydroquinolinonyl, isoquinolinyl, isoquinolinonyl,dihydroisoquinonyl or oxo-dihydroisoquinonyl, each optionallysubstituted with one to three substituents independently selected fromfluoro, chloro, methyl, methoxy, amino, methylamino, dimethylamino,amido or cyano; or a pharmaceutically acceptable salt thereof.

Yet another embodiment of the present invention is the compound of theimmediately preceding embodiment wherein R¹ is isopropyl or t-butyl; R²is hydrogen and R⁴ is hydrogen; or a pharmaceutically acceptable saltthereof. Still another embodiment of the present invention is thecompound of the immediately preceding embodiment wherein A¹ is phenyl,pyridinyl, indazolyl, indolyl, benzoimidazolyl, pyrrolopyridinyl orpyrrolopyrimidinyl; each optionally substituted with one methyl,methoxy, methylamino or dimethylamino; or a pharmaceutically acceptablesalt thereof. Another embodiment of the present invention is thecompound of either of the two immediately preceding embodiments whereinA² is phenyl substituted with CO₂H or tetrazolyl; and L is a directbond; or a pharmaceutically acceptable salt thereof.

Another embodiment of the present invention is the compound of theimmediately preceding embodiment wherein A¹ is phenyl, indolyl orbenzoimidazolyl optionally substituted with methyl, or pyridinyloptionally substituted with methylamino or dimethylamino; or apharmaceutically acceptable salt thereof.

Another embodiment of the present invention is a compound selected from:4-((4-(1-Tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)phenoxy)methyl)benzoicacid; 3-(4-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)-6-methoxypyridin-2-yl)benzoicacid;3-(4-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)-6-oxo-1,6-dihydropyridin-2-yl)benzoicacid;3-{5-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-(methylamino)pyridin-2-yl}benzoicacid;3-{5-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-(methylamino)pyridin-2-yl}benzoic acid;4′-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]biphenyl-3-carboxylicacid;4′-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]biphenyl-3-carboxylicacid;4-{5-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-(methylamino)pyridin-2-yl}benzoicacid;4-{4-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-methoxypyridin-2-yl}benzoicacid;3-{4-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-methoxypyridin-2-yl}benzoicacid;4-{4-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-methoxypyridin-2-yl}benzoicacid;3-{4-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-methoxypyridin-2-yl}benzoicacid;4-{5-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-(methylamino)pyridin-2-yl}benzoicacid;4-{5-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-(ethylamino)pyridin-2-yl}benzoicacid;4-{6-(ethylamino)-5-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]pyridin-2-yl}benzoicacid;3-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-indol-4-yl}benzoicacid;4-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-indol-4-yl}benzoicacid;3-{2-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-indol-4-yl}benzoicacid;4-{2-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-indol-4-yl}benzoicacid;3-{5-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-(ethylamino)pyridin-2-yl}benzoicacid;3-{6-(ethylamino)-5-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]pyridin-2-yl}benzoicacid;3-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-5-(1,3-oxazol-2-yl)benzoicacid;4-({4-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]phenoxy}methyl)benzoicacid;3-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-5-(1,3-oxazol-2-yl)benzoicacid;3-{6-(isopropylamino)-5-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]pyridin-2-yl}benzoicacid;4-{5-[(1-tert-butyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-(isopropylamino)pyridin-2-yl}benzoicacid;4-{6-(isopropylamino)-5-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]pyridin-2-yl}benzoicacid;4-{6-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-indazol-4-yl}benzoicacid;3-{4-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-oxo-1,6-dihydropyridin-2-yl}benzoicacid;4-{4-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-oxo-1,6-dihydropyridin-2-yl}benzoicacid;3-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-pyrrolo[2,3-b]pyridin-4-yl}benzoicacid;4-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-pyrrolo[2,3-b]pyridin-4-yl}benzoicacid;4-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-pyrrolo[3,2-c]pyridin-4-yl}benzoicacid;(5-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-indol-4-yl}-2-methoxyphenyl)aceticacid;3-{6-(dimethylamino)-4-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]pyridin-2-yl}benzoicacid;4-{6-(dimethylamino)-4-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]pyridin-2-yl}benzoicacid;4-{6-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-7H-pyrrolo[2,3-d]pyrimidin-4-yl}benzoicacid;3-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-pyrrolo[3,2-c]pyridin-4-yl}benzoicacid;3-{6-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-7H-pyrrolo[2,3-d]pyrimidin-4-yl}benzoicacid;4-{6-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-indol-4-yl}benzoicacid;4-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-pyrrolo[2,3-c]pyridin-4-yl}benzoicacid;3-{6-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-indol-4-yl}benzoicacid;3-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-pyrrolo[2,3-c]pyridin-4-yl}benzoicacid;3-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-indol-6-yl}benzoicacid;4-{5-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-[(2,2,2-trifluoroethyl)amino]pyridin-2-yl}benzoicacid;3-{5-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-(methylamino)pyridin-3-yl}benzoicacid;4-{5-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-6-(methylamino)pyridin-3-yl}benzoicacid;3-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-benzimidazol-4-yl}benzoicacid;4-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-benzimidazol-5-yl}benzoicacid;3-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-benzimidazol-5-yl}benzoicacid;3-(6-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)-2-methyl-1H-benzo[d]imidazol-4-yl)benzoicacid;4-(6-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)-2-methyl-1H-benzo[d]imidazol-4-yl)benzoicacid;1-isopropyl-1′-{[3′-(1H-tetrazol-5-yl)biphenyl-4-yl]carbonyl}-1,4-dihydrospiro[indazole-5,4′-piperidin]-7(6H)-one;and1-tert-butyl-1′-{[3′-(1H-tetrazol-5-yl)biphenyl-4-yl]carbonyl}-1,4-dihydrospiro[indazole-5,4′-piperidin]-7(6H)-one;or a pharmaceutically acceptable salt thereof.

Yet another embodiment of the present invention is the compound of theimmediately preceding embodiment selected from3-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-benzimidazol-4-yl}benzoicacid;4-{6-(dimethylamino)-4-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]pyridin-2-yl}benzoicacid;3-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-indol-4-yl}benzoicacid;3-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-pyrrolo[2,3-b]pyridin-4-yl}benzoicacid;4-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-benzimidazol-5-yl}benzoicacid;3-{2-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1′H-spiro[indazole-5,4′-piperidin]-1′-yl)carbonyl]-1H-benzimidazol-5-yl}benzoicacid;3-(6-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)-2-methyl-1H-benzo[d]imidazol-4-yl)benzoicacid; and4-(6-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)-2-methyl-1H-benzo[d]imidazol-4-yl)benzoicacid; or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention is a pharmaceutical compositioncomprising an amount of a compound of formula (I) as described in any ofthe embodiments; or a pharmaceutically acceptable salt thereof and apharmaceutically acceptable excipient, diluent, or carrier. Preferably,the composition comprises a therapeutically effective amount of acompound of the present invention or a pharmaceutically acceptable saltthereof. The composition may also contain at least one additionalpharmaceutical agent. Preferred agents include anti-diabetic agentsand/or anti-obesity agents (described herein below).

In yet another aspect of the present invention is a method for treatinga disease, condition, or disorder mediated by the inhibition ofacetyl-CoA carboxylase enzyme(s) in a mammal that includes the step ofadministering to a mammal, preferably a human, in need of such treatmenta therapeutically effective amount of a compound of the presentinvention or a pharmaceutically acceptable salt thereof, or apharmaceutical composition thereof.

Diseases, disorders, or conditions mediated by inhibitors of acetyl-CoAcarboxylases include Type II diabetes and diabetes-related diseases,such as nonalcoholic fatty liver disease (NAFLD), hepatic insulinresistance, hyperglycemia, metabolic syndrome, impaired glucosetolerance, diabetic neuropathy, diabetic nephropathy, diabeticretinopathy, obesity, dyslipidemia, hypertension, hyperinsulinemia, andinsulin resistance syndrome. Preferred diseases, disorders, orconditions include Type II diabetes, nonalcoholic fatty liver disease(NAFLD), hepatic insulin resistance, hyperglycemia, impaired glucosetolerance, obesity, and insulin resistance syndrome. More preferred areType II diabetes, nonalcoholic fatty liver disease (NAFLD), hepaticinsulin resistance, hyperglycemia, and obesity. Most preferred is TypeII diabetes.

A preferred embodiment is a method for treating, (e.g. delaying theprogression or onset) of Type 2 diabetes and diabetes-related disordersin animals comprising the step of administering to an animal in need ofsuch treatment a therapeutically effective amount of a compound of thepresent invention or a pharmaceutically acceptable salt thereof or acomposition thereof.

Another preferred embodiment is a method for treating obesity andobesity-related disorders in animals comprising the step ofadministering to an animal in need of such treatment a therapeuticallyeffective amount of a compound of the present invention or apharmaceutically acceptable salt thereof or a composition thereof.

Yet another preferred embodiment is a method for treating nonalcoholicfatty liver disease (NAFLD) or hepatic insulin resistance in animalscomprising the step of administering to an animal in need of suchtreatment a therapeutically effective amount of a compound of thepresent invention or a pharmaceutically acceptable salt thereof or acomposition thereof.

Compounds of the present invention may be administered in combinationwith other pharmaceutical agents (in particular, anti-obesity andanti-diabetic agents described herein below). The combination therapymay be administered as (a) a single pharmaceutical composition whichcomprises a compound of the present invention or a pharmaceuticallyacceptable salt thereof, at least one additional pharmaceutical agentdescribed herein and a pharmaceutically acceptable excipient, diluent,or carrier; or (b) two separate pharmaceutical compositions comprising(i) a first composition comprising a compound of the present inventionor a pharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable excipient, diluent, or carrier, and (ii) a second compositioncomprising at least one additional pharmaceutical agent described hereinand a pharmaceutically acceptable excipient, diluent, or carrier. Thepharmaceutical compositions may be administered simultaneously orsequentially and in any order.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 provides a sequence of recombinant human ACC1 (SEQ. ID NO. 1)that can be employed in the Transcreener in vitro assay.

FIG. 2 provides a sequence of recombinant human ACC2 (SEQ. ID NO. 2)that can be employed in the Transcreener in vitro assay.

DETAILED DESCRIPTION OF THE INVENTION Definitions

The phrase “therapeutically effective amount” means an amount of acompound of the present invention or a pharmaceutically acceptable saltthereof that: (i) treats or prevents the particular disease, condition,or disorder, (ii) attenuates, ameliorates, or eliminates one or moresymptoms of the particular disease, condition, or disorder, or (iii)prevents or delays the onset of one or more symptoms of the particulardisease, condition, or disorder described herein.

The term “animal” refers to humans (male or female), companion animals(e.g., dogs, cats and horses), food-source animals, zoo animals, marineanimals, birds and other similar animal species. “Edible animals” refersto food-source animals such as cows, pigs, sheep and poultry.

The phrase “pharmaceutically acceptable” indicates that the substance orcomposition must be compatible chemically and/or toxicologically, withthe other ingredients comprising a formulation, and/or the mammal beingtreated therewith.

The terms “treating”, “treat”, or “treatment” embrace both preventative,i.e., prophylactic, and palliative treatment.

The terms “modulated” or “modulating”, or “modulate(s)”, as used herein,unless otherwise indicated, refers to the inhibition of the Acetyl-CoAcarboxylases (ACC) enzyme(s) with compounds of the present invention.

The terms “mediated” or “mediating” or “mediate(s)”, as used herein,unless otherwise indicated, refers to the (i) treatment or preventionthe particular disease, condition, or disorder, (ii) attenuation,amelioration, or elimination of one or more symptoms of the particulardisease, condition, or disorder, or (iii) prevention or delay of theonset of one or more symptoms of the particular disease, condition, ordisorder described herein, by inhibiting the Acetyl-CoA carboxylases(ACC) enzyme(s).

The term “compounds of the present invention” (unless specificallyidentified otherwise) refer to compounds of Formula (I) and anypharmaceutically acceptable salts of the compounds, as well as, allstereoisomers (including diastereoisomers and enantiomers), tautomers,conformational isomers, and isotopically labeled compounds. Hydrates andsolvates of the compounds of the present invention are consideredcompositions of the present invention, wherein the compound is inassociation with water or solvent, respectively.

The terms “(C₁-C₆)alkyl” and “(C₁-C₃)alkyl” are alkyl groups of thespecified number of carbons, from one to six or one to three carbons,respectively, which can be either straight chain or branched. Forexample, the term “(C₁-C₃)alkyl” has from one to three carbons andconsists of methyl, ethyl, n-propyl and isopropyl.

The term “(C₃-C₇)cycloalkyl” means a cycloalkyl group with three toseven carbon atoms and consists of cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and cycloheptyl. The term “halo” means fluoro, chloro, bromoor iodo. The term “(C₆-C₁₀)aryl” means an aromatic carbocyclic groupconsisting of six to ten carbon atoms such as phenyl or naphthyl.

The term “5 to 12 membered heteroaryl” means a five to twelve memberedaromatic group which contains at least one heteroatom selected fromnitrogen, oxygen and sulfur. As used herein the point of attachment ofthe “5 to 12 membered heteroaryl” group is on a carbon atom of thatgroup. The “5 to 12 membered heteroaryl” group can be either monocyclicor bicyclic. Preferred embodiments of monocyclic heteroaryls include,but are not limited to, pyrazolyl, imidazolyl, triazolyl, pyridinyl, andpyrimidinyl. Preferred embodiments of bicyclic heteroaryls include, butare not limited to, radicals of the following ring systems:

The term “8 to 12 membered fused heterocyclicaryl” means an 8 to 12membered ring system in which a non-aromatic heterocyclic ring is fusedto an aryl ring. As used herein the point of attachment of the “8 to 12membered fused heterocyclicaryl” group is on a carbon atom of thatgroup. A preferred embodiment includes radicals of ring systems such as:

Compounds of the present invention may be synthesized by syntheticroutes that include processes analogous to those well-known in thechemical arts, particularly in light of the description containedherein. The starting materials are generally available from commercialsources such as Aldrich Chemicals (Milwaukee, Wis.) or are readilyprepared using methods well known to those skilled in the art (e.g.,prepared by methods generally described in Louis F. Fieser and MaryFieser, Reagents for Organic Synthesis, v. 1-19, Wiley, New York(1967-1999 ed.), or Beilsteins Handbuch der organischen Chemie, 4, Aufl.ed. Springer-Verlag, Berlin, including supplements (also available viathe Beilstein online database)).

For illustrative purposes, the reaction schemes depicted below providepotential routes for synthesizing the compounds of the present inventionas well as key intermediates. For a more detailed description of theindividual reaction steps, see the Examples section below. Those skilledin the art will appreciate that other synthetic routes may be used tosynthesize the inventive compounds. Although specific starting materialsand reagents are depicted in the schemes and discussed below, otherstarting materials and reagents can be easily substituted to provide avariety of derivatives and/or reaction conditions. In addition, many ofthe compounds prepared by the methods described below can be furthermodified in light of this disclosure using conventional chemistry wellknown to those skilled in the art.

In the preparation of compounds of the present invention, protection ofremote functionality (e.g., primary or secondary amine) of intermediatesmay be necessary. The need for such protection will vary depending onthe nature of the remote functionality and the conditions of thepreparation methods. Suitable amino-protecting groups (NH-Pg) includeacetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBz)and 9-fluorenylmethyleneoxycarbonyl (Fmoc). Similarly, a“hydroxy-protecting group” refers to a substituent of a hydroxy groupthat blocks or protects the hydroxy functionality. Suitablehydroxyl-protecting groups (O-Pg) include for example, allyl, acetyl,silyl, benzyl, para-methoxybenzyl, trityl, and the like. The need forsuch protection is readily determined by one skilled in the art. For ageneral description of protecting groups and their use, see T. W.Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, NewYork, 1991.

The following reaction schemes, Reaction Scheme I through ReactionScheme provide representative procedures that are used to preparecompounds of formula (I). It is to be understood that these reactionschemes are to be construed in a non-limiting manner and that reasonablevariations of the depicted methods can be used to prepare compounds offormula (I).

Reaction Scheme I provides three synthetic routes from penultimateintermediates to compounds of formula (I). In Equation 1 the compound ofFormula (II) is reacted with A^(2′)-L-A¹-C(O)Lg, wherein Lg is anappropriate leaving group such as hydroxy or halide, to provide thecompound of Formula (I). For example, the compound (I) may be formedusing a standard peptide coupling reaction with the desired carboxylicacid (A^(2′)-L-A¹-CO₂H, wherein A^(2′) represents wither A² itself or aprotected version of A² which can be deprotected to provide A²). Forexample, the spiropiperidine intermediate (II) and carboxylic acid(A^(2′)-L-A¹-CO₂H) may be coupled by forming an activated carboxylicacid ester, such as by contacting the carboxylic acid (A^(2′)-L-A¹-CO₂H)with a peptide coupling reagent, such asO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU) or1-Ethyl-3-(3-dimethyllaminopropyl)carbodiimide hydrochloride (EDC.HCl),in the presence or absence of an activating agent, such ashydroxybenzotriazole (HOBt) and in the presence of a suitable base, suchas N,N-diisopropylethylamine (DIEA), triethylamine or N-methylmorpholine(NMM), in a suitable solvent such as THF and/or DMF or dichloromethaneand then contacting the activated carboxylic acid ester with thespiropiperidine derivative (IIa) to form a compound of Formula (I). Thereaction can typically carried out at 0° C. to 90° C. for a period of 1to 24 hours.

Alternatively, compounds of Formula (I) can be formed by firstconverting the carboxylic acid (A^(2′)-L-A¹-CO₂H) to an acid chloride(A^(2′)-L-A¹-COCl), such as by reacting with thionyl chloride, and thenreacting the acid chloride with the spiropiperidiene derivative (IIa) inthe presence of an appropriate base such as triethylamine in anappropriate solvent such as dichloromethane to form a compound ofFormula (I). Still another alternative method entails treating thecarboxylic acid (A^(2′)-L-A¹-CO₂H) with 2-chloro-4,6-dimethoxytriazinein the presence of a suitable base, such as N-methylmorpholine in asuitable solvent such as THF and/or DMF. To the activated ester is addeda solution of the spiropiperidine derivative (IIa) and base, such asN-methylmorpholine, in a suitable solvent, such as THF and/or DMF whichthe provides the compound of formula (I).

The second and third reactions depicted in Reaction Scheme I depict thepreparation of the compound of Formula (I) using a Suzuki-type couplingreaction. The Suzuki-type coupling reactions can be carried outaccording to methods known to those skilled in the art such as thosedescribed in Miyaura, N.; Suzuki, A. Chem. Rev. 1995, 95, 2457-2483. InEquation 2 of Scheme I the compound of Formula (II′) in which Lgrepresents an appropriate leaving group such as triflate, chloro, bromoor iodo is reacted with an appropriately substituted boronate,A^(2′)-B(OR)₂. The reaction is typically carried out in the presence ofa palladium catalyst and a base in an appropriate solvent. The boronatecan be either in the form of a boronic acid or a boronic ester. InEquation 3 of Scheme I the boronate compound of Formula (II″) is reactedwith an appropriately substituted compound A^(2′)-Lg in which Lgrepresents an appropriate leaving group such as triflate, chloro, bromoor iodo. It is to be appreciated that these reactions can be carried outwhere the A¹ and A² moieties in the compounds of formulae (II′) and(II″) may contain a protected carboxylic acid group which cansubsequently be deprotected to provide an acid group in the compound offormula (I).

Reaction Scheme IA outlines the general procedures one could use toprovide compounds of the present invention having Formula (Ia) which arecompounds of Formula (I) in which R² and each R³ are each hydrogen and Zis CH₂. The protected spiropiperidine derivative (VIIIa) may be formedby treating the appropriately protected piperidine aldehyde (Xa) withmethyl vinyl ketone (IXa). The group Pg represents an appropriate amineprotecting group and is preferably N-tert-butoxycarbonyl (BOC) orcarbobenzyloxy (Cbz). This reaction can be carried out in the presenceof ethanolic potassium hydroxide according to a procedure analogous tothat described by Roy, S. et al., Chem. Eur. J. 2006, 12, 3777-3788 at3786. Alternatively, the reaction can be carried out in the presence ofpara-toluenesulfonic acid (pTSA) in refluxing benzene to provide thedesired product (VIIIa). The spiropiperidine derivative (VIIIa) can thenbe reacted with tris-(N,N-dimethylamino) methane in refluxing toluene toprovide the eneamine functionalized spiropiperidine derivative (VIIa).Compound (VIIa) is then reacted with an appropriate hydrazine derivativeR¹NHNH² in the presence of acetic acid in refluxing ethanol to providethe desired cyclized compound of formula (VIa) (see Murali Dhar, T. G.et al. Bioorg. Med. Chem. Lett. 2007, 17, 5019-5024 at 5020. Thecompound of formula (VIa) can then be treated with N-bromosuccinimide(NBS) in the presence of water in THF to provide the corresponding bromohydroxy derivative of formula (Va). The bromo hydroxy derivative (Va) isthen oxidized with Jones reagent in a method analogous to that providedin Wolinsky, J. et al., J. Org. Chem. 1978, 43(5), 875-881 at 876, 879to provide the □-bromo keto derivative of formula (IVa). The compound offormula (IVa) can then be debrominated using conventional methods suchas treatment with zinc and acetic acid or alternatively, zinc in thepresence of aqueous ammonium chloride to provide the compound of formula(IIIa).

The compound of formula (IIIa) can then be deprotected to provide thefree spiropiperidine derivative of formula (IIa) using standard methodswhich depend on which protecting group Pg has been employed. Forexample, when Pg represents tert-butyloxycarbonyl (BOC) standard strongacid deprotection conditions such as 4N hydrochloric acid in dioxane ortrifluoroacetic acid in an appropriate solvent such as dichloromethanecan be used to remove the BOC group. When Pg represents carbobenzyloxy(Cbz), hydrogenation over palladium on carbon in ethanol or treatmentwith ammonium formate in the presence of palladium on carbon in ethanolcan be employed to carry out the deprotection.

The spiropiperidine derivative of Formula (IIa) can then be acylated byemploying standard methods to provide the compound of Formula (Ia). Forexample, the compound (la) may then be formed using a standard peptidecoupling reaction with the desired carboxylic acid (A^(2′)-L-A¹-CO₂H,wherein A^(2′) represents either A² itself or a protected version of A²which can be deprotected to provide A²). For example, Thespiropiperidine intermediate (IIa) and carboxylic acid(A^(2′)-L-A¹-CO₂H) may be coupled by forming an activated carboxylicacid ester, such as by contacting the carboxylic acid (A^(2′)-L-A¹-CO₂H)with a peptide coupling reagent, such asO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU) or 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC.HCl), in the presence or absence of anactivating agent, such as hydroxybenzotriazole (HOBt) and in thepresence of a suitable base, such as N,N-diisopropylethylamine (DIEA),triethylamine or N-methylmorpholine (NMM), in a suitable solvent such asTHF and/or DMF or dichloromethane and then contacting the activatedcarboxylic acid ester with the spiropiperidine derivative (IIa) to forma compound of Formula (Ia).

Alternatively, compounds of Formula (Ia) can be formed by firstconverting the carboxylic acid (A^(2′)-L-A¹-CO₂H) to an acid chloride(A^(2′)-L-A¹-COCl), such as by reacting with thionyl chloride, and thenreacting the acid chloride with the spiropiperidiene derivative (IIa) inthe presence of an appropriate base such as triethylamine in anappropriate solvent such as dichloromethane to form a compound ofFormula (Ia). Still another alternative method entails treating thecarboxylic acid (A^(2′)-L-A¹-CO₂H) with 2-chloro-4,6-dimethoxytriazinein the presence of a suitable base, such as N-methylmorpholine in asuitable solvent such as THF and/or DMF. To the activated ester is addeda solution of the spiropiperidine derivative (IIa) and base, such asN-methylmorpholine, in a suitable solvent, such as THF and/or DMF whichthe provides the compound of formula (Ia).

Reaction Scheme II provides an alternative synthesis of compounds offormula (Ia) starting from the intermediate of formula (VIa). Thecompound of formula (VIa) is treated with N-bromosuccinimide (NBS) inthe presence of methanol in THF (Nishimura, T. et al. Org. Lett. 2008,10(18), 4057-4060 at 4059) to provide the methoxy bromo spiropiperidinederivative of formula (Vb). Base induced elimination of compound (Vb) bytreatment with a strong base such as potassium tert-butoxide in THFprovides the compound of formula (IVb) which is then treated with astrong acid such as 2N hydrochloric acid in THF to provide the compoundof formula (IIIa). Compound (IIIa) can then be deprotected and acylatedas described previously in Reaction Scheme I to provide compounds offormula (Ia).

Reaction Scheme III provides a synthesis of compounds of formula (Ib)which are compounds of formula (I) in which R² is bromo and each R³ ishydrogen. The compound of formula (VIa) is reacted with approximatelytwo equivalents of N-bromosuccinimide in the presence of methanol toprovide the dibromo methoxy spiropiperidine derivative of formula (Vc).Compound (Vc) is then subjected to elimination conditions by treatmentwith a strong base such as potassium tert-butoxide in an appropriatesolvent to provide the compound of formula (IVc). Treatment of thecompound of formula (IVc) with strong acid such as 2N hydrochloric acidprovides the compound of formula (IIIb). Deprotection of compound (IIIb)to provide compound (IIb) followed by acylation to provide the compoundof formula (Ib) can be carried out as described previously for ReactionScheme I.

Reaction Scheme IV depicts the preparation of certain other compoundswithin formula (I) from certain of the intermediates previouslydepicted. The first reaction in Scheme IV shows introduction of a methylgroup at the R² position by reacting the bromo spiropiperidinederivative of formula (IIIb) with trimethoxyborate in the presence of anappropriate palladium catalyst, such as palladium tetrakistriphenylphosphine in the presence of potassium carbonate and water toprovide (IIIc). Other alkyl groups can be introduced at the R² positionin an analogous manner. The compound of formula (IIIc) can then bedeprotected and acylated as previously described. The second reaction inReaction Scheme IV depicts introduction of a cyano group at the R²position. The bromo spiropiperidine compound (IIIb) is reacted with zinccyanide in the presence of zinc and an appropriate palladium catalyst toprovide (IIId) which can then be deprotected and acylated to provide acompound of formula (Id). The third reaction in Scheme IV depictsintroduction of an appropriate group at the R³ position of the compound(IIIe). The compound (IIIe) is deprotonated with a strong base, such aslithium hexamethyldisilazide (LHMDS) under appropriate anhydrousconditions in an appropriate solvent, preferably at low temperature. Theenolate thus formed is then reacted with an appropriate electrophileR³Lg wherein Lg represents an appropriate leaving group (such as ahalide when R³Lg is an alkyl halide) to provide (IIIf) wherein R³ is anappropriate group such as an alkyl group. The deprotonation of (IIIf)and reaction with another R³Lg can then be carried out again if desiredto prepare a di-R³ (IIIf) compound wherein the R³ groups may be the sameor different. The compound of formula (IIIf) can then be deprotected andacylated as previously described to provide the compound of formula(Ie).

Reaction Scheme VI provides the synthesis of compounds within Formula(I) wherein R¹ is isopropyl, R² is hydrogen, each R³ is hydrogen and Zis oxygen. 1-(1-(4-methoxy benzyl)-4-hydroxy-1H-pyrazol-3-yl)ethanone isreacted with 1-benzyl piperidin-4-one in refluxing methanol in thepresence of pyrrolidine to provide the diprotected spiro-compound(IIIg). The para-methoxybenzyl group of (IIIg) is then removed upontreatment with trifluoroacetic acid in dichloroethane at an elevatedtemperature, such as 90° C., to provide the benzyl protected N-1 (H)pyrazole derivative (IIh). This benzyl protected N-1(H) pyrazolederivative compound is then subjected to Mitsonubo coupling conditionsusing isopropanol in the presence of Di-t-butylazodicarboxylate (DBAD)and triphenylphosphine in tetrahydrofuran to provide the correspondingbenzyl protected N-1-isopropyl compound (IIIi). The benzyl protectedN-1-isopropyl compound can then be debenzylated upon treatment withα-chloroethyl chloroformate (ACE-Cl) and methanol to provide thecorresponding free spiropiperidine derivative (IIc). The freespiropiperidine derivative (IIc) can then be acylated as previouslydescribed to provide the compounds of Formula (If).

Reaction Scheme VII depicts the hydrolysis of a protected acidintermediate of Formula (Ij) to provide the acid bearing compound ofFormula (Ig). For example, the compound of Formula (Ij) where Rrepresents an appropriate acid protecting group such as t-butyl orpara-methoxybenzyl can be treated with a strong acid such astrifluoroacetic acid or hydrochloric acid in an appropriate solvent suchas dichloromethane to provide the compound of Formula (Ig). In thisReaction Scheme the acid group is shown as appended onto A^(2′) and theacid taken together with A^(2′) represent the group A² in the compoundof Formula (I). It is to be appreciated that the acid group may also bepart of A¹ in a like manner.

Reaction Scheme VIII provides methods useful for preparing certainintermediates useful in the preparation of compounds of Formula (I).Equation 1 of Reaction Scheme VIII provides a Suzuki-type couplingbetween an appropriate acid derivative X-A¹-CO₂R^(a) with an appropriateboronate (RO)₂B-A^(2′)-CO₂R^(b) wherein R^(a) and R^(b) aredifferentially protected or one of R^(a) and R^(b) is hydrogen, X is ahalide or sulfonate such as triflate and R is hydrogen or an alkyl suchas methyl. Equation 2 of Reaction Scheme VIII provides anotherSuzuki-type coupling between X-A¹-CO₂R^(a) with an appropriate boronate(RO)₂B-A^(2′)-tetrazolyl. The Suzuki-type coupling can be carried out asdescribed previously in Reaction Scheme I. The final intermediatecompound of Equation 1 and 2 wherein R^(a) is hydrogen can then be usedin acylation type reactions with a compound of Formula (II) as describedin Equation 1 of Reaction Scheme I.

Reaction Scheme IX provides another method useful for preparing certainintermediates useful for preparing compounds of Formula (I). Equation 1of Reaction Scheme IX depicts the reaction of 2,6-dichloronicotinic acidwith an appropriately substituted boronate (where R is hydrogen or alkylsuch as methyl and R^(b) is typically an acid protecting group such ast-butyl) under Suzuki-type coupling conditions to provide the2-chloro-6-substituted nicotinic acid. The 2-chloro-6-substitutednicotinic acid can then be reacted with an appropriate nucleophile HY-R′(wherein R′ is typically alkyl optionally substituted with halo, R″ istypically alkyl such as methyl, ethyl, propyl or isopropyl) to providethe disubstituted nicotinic acid derivative. Alternatively, the reactioncan be carried out by first reacting it with the nucleophile HY-R′followed by the Suzuki-type coupling with the boronate as describedabove. The disubstituted nicotinic acid derivative can then be employedin acylation reactions with compounds of Formula (II) followed bydeprotection as necessary as described in Reaction Scheme I to providecompounds of Formula (I) wherein A¹ is the substituted pyridine moietyas shown. Equation 2 of Reaction Scheme IX depicts reacting5-bromo-6-chloronicotinic acid with an appropriate nucleophile HY-R′ toprovide the 5-bromo-6-substituted nicotinic acid derivative which isthen reacted with an appropriate boronate under Suzuli-type couplingconditions to provide the 5,6-disubstituted nicotinic acid derivative.The 5,6-disubstituted nicotinic acid derivative can then be employed inacylation reactions with compounds of Formula (II) followed bydeprotection as necessary as previously described in Reaction Scheme Ito provide compounds of Formula (I) wherein A¹ is the substitutedpyridine moiety as shown.

The compounds of the present invention may be isolated and used per seor in the form of their pharmaceutically acceptable salts. In accordancewith the present invention, compounds with multiple basic nitrogen atomscan form salts with varying number of equivalents (“eq.”) of acid. Itwill be understood by practitioners that all such salts are within thescope of the present invention.

Pharmaceutically acceptable salts, as used herein in relation tocompounds of the present invention, include pharmaceutically acceptableinorganic and organic salts of said compound. These salts can beprepared in situ during the final isolation and purification of acompound, or by separately reacting the compound thereof, with asuitable organic or inorganic acid and isolating the salt thus formed.Representative salts include, but are not limited to, the hydrobromide,hydrochloride, hydroiodide, sulfate, bisulfate, nitrate, acetate,trifluoroacetate, oxalate, besylate, palmitate, pamoate, malonate,stearate, laurate, malate, borate, benzoate, lactate, phosphate,hexafluorophosphate, benzene sulfonate, tosylate, formate, citrate,maleate, fumarate, succinate, tartrate, naphthylate, mesylate,glucoheptonate, lactobionate and laurylsulphonate salts, and the like.These may also include cations based on the alkali and alkaline earthmetals, such as sodium, lithium, potassium, calcium, magnesium, and thelike, as well as non-toxic ammonium, quaternary ammonium, and aminecations including, but not limited to, ammonium, tetramethylammonium,tetraethylammonium, methylammonium, dimethylammonium, trimethylammonium,triethylammonium, ethylammonium, and the like. For additional examplessee, for example, Berge, et al., J. Pharm. Sci., 66, 1-19 (1977).

Certain compounds of the present invention may exist in more than onecrystal form. Polymorphs of compounds of Formula (I) and salts thereof(including solvates and hydrates) form part of this invention and may beprepared by crystallization of a compound of the present invention underdifferent conditions. For example, using different solvents or differentsolvent mixtures for recrystallization; crystallization at differenttemperatures; various modes of cooling, ranging from very fast to veryslow cooling during crystallization. Polymorphs may also be obtained byheating or melting a compound of the present invention followed bygradual or fast cooling. The presence of polymorphs may be determined bysolid probe nuclear magnetic resonance (NMR) spectroscopy, infrared (IR)spectroscopy, differential scanning calorimetry, powder X-raydiffraction or such other techniques.

This invention also includes isotopically-labeled compounds, which areidentical to those described by Formula (1), but for the fact that oneor more atoms are replaced by an atom having an atomic mass or massnumber different from the atomic mass or mass number usually found innature. Examples of isotopes that can be incorporated into compounds ofthe invention include isotopes of hydrogen, carbon, nitrogen, oxygen,sulfur and fluorine, such as ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³⁵S, ³⁶Cl,¹²⁵I, ¹²⁹I, and ¹⁸F respectively. Certain isotopically-labeled compoundsof the present invention, for example those into which radioactiveisotopes such as ³H and ¹⁴C are incorporated, are useful in drug and/orsubstrate tissue distribution assays. Tritiated (i.e., ³H), andcarbon-14 (i.e., ¹⁴C), isotopes are particularly preferred for theirease of preparation and detectability. Further, substitution withheavier isotopes such as deuterium (i.e., ²H), can afford certaintherapeutic advantages resulting from greater metabolic stability, forexample increased in vivo half-life or reduced dosage requirements and,hence, may be preferred in some circumstances. Isotopically labeledcompounds of the present invention can generally be prepared by carryingout the procedures disclosed in the schemes and/or in the Examplesbelow, by substituting a readily available isotopically labeled reagentfor a non-isotopically labeled reagent.

The compounds of the present invention may contain stereogenic centers.These compounds may exist as mixtures of enantiomers or as pureenantiomers. Wherein a compound includes a stereogenic center, thecompounds may be resolved into the pure enantiomers by methods known tothose skilled in the art, for example by formation of diastereoisomericsalts which may be separated, for example, by crystallization; formationof stereoisomeric derivatives or complexes which may be separated, forexample, by crystallization, gas-liquid or liquid chromatography;selective reaction of one enantiomer with an enantiomer-specificreagent, for example enzymatic esterification; or gas-liquid or liquidchromatography in a chiral environment, for example on a chiral supportfor example silica with a bound chiral ligand or in the presence of achiral solvent. It will be appreciated that where the desiredstereoisomer is converted into another chemical entity by one of theseparation procedures described above, a further step is required toliberate the desired enantiomeric form. Alternatively, the specificstereoisomers may be synthesized by using an optically active startingmaterial, by asymmetric synthesis using optically active reagents,substrates, catalysts or solvents, or by converting one stereoisomerinto the other by asymmetric transformation.

Certain compounds of the present invention may exist in different stableconformational forms which may be separable. Torsional asymmetry due torestricted rotation about an asymmetric single bond, for example becauseof steric hindrance or ring strain, may permit separation of differentconformers. The compounds of the present invention further include eachconformational isomer of compounds of Formula (1) and mixtures thereof.

Compounds of the present invention are useful for treating diseases,conditions and/or disorders modulated by the inhibition of theacetyl-CoA carboxylases enzyme(s) (in particular, ACC1 and ACC2);therefore, another embodiment of the present invention is apharmaceutical composition comprising a therapeutically effective amountof a compound of the present invention and a pharmaceutically acceptableexcipient, diluent or carrier. The compounds of the present invention(including the compositions and processes used therein) may also be usedin the manufacture of a medicament for the therapeutic applicationsdescribed herein.

A typical formulation is prepared by mixing a compound of the presentinvention and a carrier, diluent or excipient. Suitable carriers,diluents and excipients are well known to those skilled in the art andinclude materials such as carbohydrates, waxes, water soluble and/orswellable polymers, hydrophilic or hydrophobic materials, gelatin, oils,solvents, water, and the like. The particular carrier, diluent orexcipient used will depend upon the means and purpose for which thecompound of the present invention is being applied. Solvents aregenerally selected based on solvents recognized by persons skilled inthe art as safe (GRAS) to be administered to a mammal. In general, safesolvents are non-toxic aqueous solvents such as water and othernon-toxic solvents that are soluble or miscible in water. Suitableaqueous solvents include water, ethanol, propylene glycol, polyethyleneglycols (e.g., PEG400, PEG300), etc. and mixtures thereof. Theformulations may also include one or more buffers, stabilizing agents,surfactants, wetting agents, lubricating agents, emulsifiers, suspendingagents, preservatives, antioxidants, opaquing agents, glidants,processing aids, colorants, sweeteners, perfuming agents, flavoringagents and other known additives to provide an elegant presentation ofthe drug (i.e., a compound of the present invention or pharmaceuticalcomposition thereof) or aid in the manufacturing of the pharmaceuticalproduct (i.e., medicament).

The formulations may be prepared using conventional dissolution andmixing procedures. For example, the bulk drug substance (i.e., compoundof the present invention or stabilized form of the compound (e.g.,complex with a cyclodextrin derivative or other known complexationagent)) is dissolved in a suitable solvent in the presence of one ormore of the excipients described above. The dissolution rate of poorlywater-soluble compounds may be enhanced by the use of a spray-drieddispersion, such as those described by Takeuchi, H., et al. in“Enhancement of the dissolution rate of a poorly water-soluble drug(tolbutamide) by a spray-drying solvent depostion method anddisintegrants” J. Pharm. Pharmacol., 39, 769-773 (1987); and EP0901786B1 (US2002/009494), incorporated herein by reference. The compound ofthe present invention is typically formulated into pharmaceutical dosageforms to provide an easily controllable dosage of the drug and to givethe patient an elegant and easily handleable product.

The pharmaceutical compositions also include solvates and hydrates ofthe compounds of the present invention. The term “solvate” refers to amolecular complex of a compound represented by Formula (I) (includingpharmaceutically acceptable salts thereof) with one or more solventmolecules. Such solvent molecules are those commonly used in thepharmaceutical art, which are known to be innocuous to the recipient,e.g., water, ethanol, ethylene glycol, and the like, The term “hydrate”refers to the complex where the solvent molecule is water. The solvatesand/or hydrates preferably exist in crystalline form. Other solvents maybe used as intermediate solvates in the preparation of more desirablesolvates, such as methanol, methyl t-butyl ether, ethyl acetate, methylacetate, (S)-propylene glycol, (R)-propylene glycol, 1,4-butyne-diol,and the like.

The pharmaceutical composition (or formulation) for application may bepackaged in a variety of ways depending upon the method used foradministering the drug. Generally, an article for distribution includesa container having deposited therein the pharmaceutical formulation inan appropriate form. Suitable containers are well-known to those skilledin the art and include materials such as bottles (plastic and glass),sachets, ampoules, plastic bags, metal cylinders, and the like. Thecontainer may also include a tamper-proof assemblage to preventindiscreet access to the contents of the package. In addition, thecontainer has deposited thereon a label that describes the contents ofthe container. The label may also include appropriate warnings.

The present invention further provides a method of treating diseases,conditions and/or disorders modulated by the inhibition of theacetyl-CoA carboxylases enzyme(s) in an animal that includesadministering to an animal in need of such treatment a therapeuticallyeffective amount of a compound of the present invention or apharmaceutical composition comprising an effective amount of a compoundof the present invention and a pharmaceutically acceptable excipient,diluent, or carrier. The method is particularly useful for treatingdiseases, conditions and/or disorders that benefit from the inhibitionof acetyl-CoA carboxylases enzyme(s).

One aspect of the present invention is the treatment of obesity, andobesity-related disorders (e.g., overweight, weight gain, or weightmaintenance).

Obesity and overweight are generally defined by body mass index (BMI),which is correlated with total body fat and estimates the relative riskof disease. BMI is calculated by weight in kilograms divided by heightin meters squared (kg/m²). Overweight is typically defined as a BMI of25-29.9 kg/m², and obesity is typically defined as a BMI of 30 kg/m².See, e.g., National Heart, Lung, and Blood Institute, ClinicalGuidelines on the Identification, Evaluation, and Treatment ofOverweight and Obesity in Adults, The Evidence Report, Washington, D.C.:U.S. Department of Health and Human Services, NIH publication no.98-4083 (1998).

Another aspect of the present invention is for the treatment or delayingthe progression or onset of diabetes or diabetes-related disordersincluding Type 1 (insulin-dependent diabetes mellitus, also referred toas “IDDM”) and Type 2 (noninsulin-dependent diabetes mellitus, alsoreferred to as “NIDDM”) diabetes, impaired glucose tolerance, insulinresistance, hyperglycemia, and diabetic complications (such asatherosclerosis, coronary heart disease, stroke, peripheral vasculardisease, nephropathy, hypertension, neuropathy, and retinopathy).

In yet another aspect of the present invention is the treatment ofobesity co-morbidities, such as metabolic syndrome. Metabolic syndromeincludes diseases, conditions or disorders such as dyslipidemia,hypertension, insulin resistance, diabetes (e.g., Type 2 diabetes),coronary artery disease and heart failure. For more detailed informationon Metabolic Syndrome, see, e.g., Zimmet, P. Z., et al., “The MetabolicSyndrome: Perhaps an Etiologic Mystery but Far From a Myth—Where Doesthe International Diabetes Federation Stand?,” Diabetes & Endocrinology,7(2), (2005); and Alberti, K. G., et al., “The Metabolic Syndrome—A NewWorldwide Definition,” Lancet, 366, 1059-62 (2005). Preferably,administration of the compounds of the present invention provides astatistically significant (p<0.05) reduction in at least onecardiovascular disease risk factor, such as lowering of plasma leptin,C-reactive protein (CRP) and/or cholesterol, as compared to a vehiclecontrol containing no drug. The administration of compounds of thepresent invention may also provide a statistically significant (p<0.05)reduction in glucose serum levels.

In yet another aspect of the invention is the treatment of nonalcoholicfatty liver disease (NAFLD) and heptic insulin resistance.

For a normal adult human having a body weight of about 100 kg, a dosagein the range of from about 0.001 mg to about 10 mg per kilogram bodyweight is typically sufficient, preferably from about 0.01 mg/kg toabout 5.0 mg/kg, more preferably from about 0.01 mg/kg to about 1 mg/kg.However, some variability in the general dosage range may be requireddepending upon the age and weight of the subject being treated, theintended route of administration, the particular compound beingadministered and the like. The determination of dosage ranges andoptimal dosages for a particular patient is well within the ability ofone of ordinary skill in the art having the benefit of the instantdisclosure. It is also noted that the compounds of the present inventioncan be used in sustained release, controlled release, and delayedrelease formulations, which forms are also well known to one of ordinaryskill in the art.

The compounds of the present invention may also be used in conjunctionwith other pharmaceutical agents for the treatment of the diseases,conditions and/or disorders described herein. Therefore, methods oftreatment that include administering compounds of the present inventionin combination with other pharmaceutical agents are also provided.Suitable pharmaceutical agents that may be used in combination with thecompounds of the present invention include anti-obesity agents(including appetite suppressants), anti-diabetic agents,anti-hyperglycemic agents, lipid lowering agents, and anti-hypertensiveagents.

Suitable anti-obesity agents include 11β-hydroxy steroid dehydrogenase-1(11β-HSD type 1) inhibitors, stearoyl-CoA desaturase-1 (SCD-1)inhibitor, MCR-4 agonists, cholecystokinin-A (CCK-A) agonists, monoaminereuptake inhibitors (such as sibutramine), sympathomimetic agents, β₃adrenergic agonists, dopamine agonists (such as bromocriptine),melanocyte-stimulating hormone analogs, 5HT2c agonists, melaninconcentrating hormone antagonists, leptin (the OB protein), leptinanalogs, leptin agonists, galanin antagonists, lipase inhibitors (suchas tetrahydrolipstatin, i.e. orlistat), anorectic agents (such as abombesin agonist), neuropeptide-Y antagonists (e.g., NPY Y5antagonists), PYY₃₋₃₆ (including analogs thereof), thyromimetic agents,dehydroepiandrosterone or an analog thereof, glucocorticoid agonists orantagonists, orexin antagonists, glucagon-like peptide-1 agonists,ciliary neurotrophic factors (such as Axokine™ available from RegeneronPharmaceuticals, Inc., Tarrytown, N.Y. and Procter & Gamble Company,Cincinnati, Ohio), human agouti-related protein (AGRP) inhibitors,ghrelin antagonists, histamine 3 antagonists or inverse agonists,neuromedin U agonists, MTP/ApoB inhibitors (e.g., gut-selective MTPinhibitors, such as dirlotapide), opioid antagonist, orexin antagonist,and the like.

Preferred anti-obesity agents for use in the combination aspects of thepresent invention include gut-selective MTP inhibitors (e.g.,dirlotapide, mitratapide and implitapide, R56918 (CAS No. 403987) andCAS No. 913541-47-6), CCKa agonists (e.g.,N-benzyl-2-[4-(1H-indol-3-ylmethyl)-5-oxo-1-phenyl-4,5-dihydro-2,3,6,10b-tetraaza-benzo[e]azulen-6-yl]-N-isopropyl-acetamidedescribed in PCT Publication No. WO 2005/116034 or US Publication No.2005-0267100 A1), 5HT2c agonists (e.g., lorcaserin), MCR4 agonist (e.g.,compounds described in U.S. Pat. No. 6,818,658), lipase inhibitor (e.g.,Cetilistat), PYY₃₋₃₆ (as used herein “PYY₃₋₃₆” includes analogs, such aspeglated PYY₃₋₃₆ e.g., those described in US Publication 2006/0178501),opioid antagonists (e.g., naltrexone), oleoyl-estrone (CAS No.180003-17-2), obinepitide (TM30338), pram lintide (Symlin®), tesofensine(NS2330), leptin, liraglutide, bromocriptine, orlistat, exenatide(Byetta®), AOD-9604 (CAS No. 221231-10-3) and sibutramine. Preferably,compounds of the present invention and combination therapies areadministered in conjunction with exercise and a sensible diet.

Suitable anti-diabetic agents include a sodium-glucose co-transporter(SGLT) inhibitor, a phosphodiesterase (PDE)-10 inhibitor, adiacylglycerol acyltransferase (DGAT) 1 or 2 inhibitor, a sulfonylurea(e.g., acetohexamide, chlorpropamide, diabinese, glibenclamide,glipizide, glyburide, glimepiride, gliclazide, glipentide, gliquidone,glisolamide, tolazamide, and tolbutamide), a meglitinide, an α-amylaseinhibitor (e.g., tendamistat, trestatin and AL-3688), an α-glucosidehydrolase inhibitor (e.g., acarbose), an α-glucosidase inhibitor (e.g.,adiposine, cam iglibose, emiglitate, miglitol, voglibose, pradimicin-Q,and salbostatin), a PPARy agonist (e.g., balaglitazone, ciglitazone,darglitazone, englitazone, isaglitazone, pioglitazone, rosiglitazone andtroglitazone), a PPAR α/γ agonist (e.g., CLX-0940, GW-1536, GW-1929,GW-2433, KRP-297, L-796449, LR-90, MK-0767 and SB-219994), a biguanide(e.g., metformin), a glucagon-like peptide 1 (GLP-1) agonist (e.g.,Byetta™, exendin-3 and exendin-4), a protein tyrosine phosphatase-1B(PTP-1B) inhibitor (e.g., trodusquemine, hyrtiosal extract, andcompounds disclosed by Zhang, S., et al., Drug Discovery Today,12(9/10), 373-381 (2007)), SIRT-1 inhibitor (e.g., reservatrol), adipeptidyl peptidease IV (DPP-IV) inhibitor (e.g., sitagliptin,vildagliptin, alogliptin and saxagliptin), an insulin secreatagogue, afatty acid oxidation inhibitor, an A2 antagonist, a c-jun amino-terminalkinase (JNK) inhibitor, insulin, an insulin mimetic, a glycogenphosphorylase inhibitor, a VPAC2 receptor agonist and a glucokinaseactivator. Preferred anti-diabetic agents are metformin, a glucagon-likepeptide 1 (GLP-1) agonist (e.g, Byetta™) and DPP-IV inhibitors (e.g.,sitagliptin, vildagliptin, alogliptin and saxagliptin).

All of the above recited U.S. patents and publications are incorporatedherein by reference.

The Examples set forth herein below are for illustrative purposes only.The compositions, methods, and various parameters reflected herein areintended only to exemplify various aspects and embodiments of theinvention, and are not intended to limit the scope of the claimedinvention in any way.

EXAMPLES

The compounds and intermediates described below were generally namedaccording to the IUPAC (International Union for Pure and AppliedChemistry) recommendations on Nomenclature of Organic Chemistry and theCAS Index rules. Unless noted otherwise, all reactants were obtainedcommercially. All of the references cited herein below are incorporatedby reference.

-   Flash chromatography was performed according to the method described    by Still et al., J. Org. Chem., 1978, 43, 2923.-   All Biotage® purifications, discussed herein, were performed using    either a 40M or 40S Biotage® column containing KP-SIL silica (40-63    μM, 60 Angstroms) (Bioatge AB; Uppsala, Sweden).-   All Combiflash® purifications, discussed herein, were performed    using a CombiFlash® Companion system (Teledyne Isco; Lincoln, Nebr.)    utilizing packed RediSep® silica columns-   Mass Spectra were recorded on a Waters (Waters Corp.; Milford,    Mass.) Micromass Platform II spectrometer. Unless otherwise    specified, mass spectra were recorded on a Waters (Milford, Mass.)    Micromass Platform II spectrometer.-   Proton NMR chemical shifts are given in parts per million downfield    from tetramethylsilane and were recorded on a Varian Unity 400 or    500 MHz (megaHertz) spectrometer (Varian Inc.; Palo Alto, Calif.).    NMR chemical shifts are given in parts per million downfield from    tetramethylsilane (for proton) or fluorotrichloromethane (for    fluorine).

The preparations described below were used in the synthesis of compoundsexemplified in the following examples.

Preparation of Intermediates and Starting Materials Intermediate 1

5-(4-(Tert-butoxycarbonyl)phenyl)-6-ethoxynicotinic acid, shown below,was prepared as follows.

Step1. 5-Bromo-6-ethoxynicotinic acid, shown below, was prepared asfollows.

A slurry of 5-bromo-6-chloronicotinic acid (240 mg, 1.0 mmol) and sodiumethoxide (138 mg, 2.0 mmol) in anhydrous ethanol (2 mL) was heated undermicrowave conditions at 100° C. for 15 min; an additional portion ofsodium ethoxide (79 mg, 1.0 mmol) was added and heating was continuedfor 1 hr. After cooling the reaction mixture was adjusted to a pH of 4with 1 N aqueous hydrochloric acid, the resulting solids collected anddried in vacuo to afford 5-bromo-6-ethoxynicotinic acid (140 mg). 1H NMR(400 MHz, DMSO-d6) d ppm 1.33 (t, J=7.02 Hz, 3H) 4.43 (q, J=7.09 Hz, 2H)8.32 (d, J=2.15 Hz, 1H) 8.64 (d, J=1.95 Hz, 1H) 13.28 (br. s., 1H);m/z=248.2 (M+1).

Step 2. The title compound, shown above, was prepared as follows: Aslurry of 5-bromo-6-ethoxynicotinic acid (60 mg, 0.24 mmol),4-tert-butoxycarbonylphenylboronic acid (70 mg, 0.32 mmol), 2 N aqueoussodium carbonate (0.37 mL, 0.73 mmol) and palladium1,1′-bis(diphenylphosphino)ferrocene dichloride (9 mg, 0.05 mmol) inp-dioxane (2 mL) were heated at 100° C. for 2 hr. An additional portionof 4-tert-butylcarboxylphenylboronic acid (70 mg, 0.32 mmol) andpalladium 1,1′-bis(diphenylphosphino)ferrocene dichloride (9 mg, 0.05mmol) were added and heating was continued for 1.5 hr. The reactionmixture was cooled, diluted into water, pH adjusted to ˜5 using 1 Naqueous hydrochloric acid. This mixture was extracted with ethyl acetate(3×), the combined organic layers washed with brine, dried overmagnesium sulfate and concentrated in vacuo to afford the title compound(100 mg), which was utilized without further purification; m/z=344.2(M+1).

Intermediate 2

2-(4-(Tert-butoxycarbonyl)phenyl)-6-(dimethylamino)isonicotinic acid,shown below, was prepared as follows.

Step 1. 2-Chloro-6-(dimethylamino)isonicotinic acid, hydrochloride salt,shown below, was prepared as follows.

2,6-Dichloroisonicotinic acid (2.00 g, 10.42 mmol) was placed in apressure tube and a solution of dimethylamine in tetrahydrofuran (26 mL,2 M, 52 mmol) added. The vessel was sealed and heated for 22 h at 80° C.The mixture was cooled to room temperature, transferred to around-bottom flask and concentrated to dryness. The resulting whitesemi-solid was taken up in 30 mL of 0.1 N sodium hydroxide solution. 1 NHCl was added dropwise with stirring to adjust the pH of the solution toca. 3.5, at which point a pale yellow solid formed. This was collectedby filtration and dried under vacuum at 45° C. overnight to provide2-chloro-6-(dimethylamino)isonicotinic acid inner salt (916 mg, 44%).Further acidification of the aqueous solution to pH 1 resulted in theformation of a bright yellow solid, which was also collected and driedunder vacuum to give 2-chloro-6-(dimethylamino) isonicotinic acidhydrochloride (1.15 g, 46%). m/z: 201+[M+H]; 199−[M−H]. For the HClsalt: ¹H NMR (400 MHz, DMSO-d₆) δ ppm 13.64 (br. s., 1H), 6.96 (d, J=1.0Hz, 1H), 6.89 (d, J=0.8 Hz, 1H), 3.06 (s, 6H), 2.53 (t, J=5.1 Hz, 1H).

For the inner salt: ¹H NMR (400 MHz, DMSO-d₆) δ ppm 6.95 (s, 1H), 6.88(s, 1H), 3.04 (s, 6H).

Step 2. 2-(4-(tert-butoxycarbonyl)phenyl)-6-(dimethylamino)isonicotinicacid, shown below, was prepared as follows.

2-Chloro-6-(dimethylamino)isonicotinic acid (450 mg, 2.24 mmol),4-(tert-butoxycarbonyl)benzene boronic acid (648 mg, 2.92 mmol),1,4-dioxane (7.5 mL) and sodium carbonate (713 mg, 6.73 mmol) dissolvedin water (3.36 mL) were placed in a flask and the mixture bubbled withnitrogen while stirring for 10 min. Palladium(II) acetate (20 mg, 0.09mmol) and 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (75 mg, 0.18mmol) were then added together and the vessel flushed with nitrogen,sealed, and heated at 90° C. for 5 h. The mixture was then cooled toroom temperature, diluted with ethyl acetate (50 mL), acidified to pH 2with 1.5 N HCl and filtered through a pad of celite. The layers wereseparated and the aqueous portion extracted with ethyl acetate (2×50mL). The combined organic portions were treated with anhydrous sodiumsulfate and decolorizing charcoal and stirred for 30 min beforefiltering. The solution was concentrated to dryness and the residuepurified by trituration with a mixture of methyl tert-butyl ether (5 mL)and heptane (100 mL). The solids were collected by filtration and driedto give 2-(4-(tert-butoxycarbonyl)phenyl)-6-(dimethylamino)isonicotinicacid (502 mg, 65%) as a pale yellow powder. m/z: 343+[M+H]; 341−[M−H];¹H NMR (400 MHz, CDCl₃) δ ppm 8.12-8.21 (m, 2H), 8.04-8.11 (m, 2H), 7.67(s, 1H), 7.16 (s, 1H), 3.23 (s, 6H), 1.64 (s, 9H).

Intermediate 3

2-(4-(Tert-butoxycarbonyl)phenyl)-6-ethoxyisonicotinic acid, shownbelow, was prepared as follows.

Sodium (65 mg, 3 mmol) was added to absolute ethanol (3 mL) under argonat RT. After completed dissolution of the sodium, the freshly preparedethoxide solution was added to 2,6-dichloronicotinic acid 1 (0.5 g, 2.6mmol). This mixture was heated in a microwave at 150° C. for 3 hrs. Themixture was concentrated to dryness to provide crude2-chloro-6-ethoxyisonicotinic acid (0.5 g), which was used withoutpurification. m/z: 202+[M+H]

The crude 2-chloro-6-ethoxyisonicotinic acid (0.5 g, ca. 2.6 mmol),4-(tert-butoxycarbonyl) benzene boronic acid (0.5 g, 2.2 mmol),potassium carbonate (0.5 g, 3.6 mmol) and water (0.1 mL) in1,2-dimethoxyethane (10 mL) were stirred and degassed for 10 min.Tetrakis(triphenylphosphine)palladium (0.1 g, 0.08 mmol) was then added.The sealed tube was then heated over night at 90° C. After cooling thereaction was passed through a plug a celite and partitioned betweendichloromethane and water. The organic solution was concentrated and thecrude residue purified by chromatography (silica gel, 10% methanol indichloromethane) to afford the title compound (97 mg, 20%). ¹H NMR (400MHz, CDCl₃) δ ppm 8.11 (2H, d), 8.08 (2H, d), 7.96 (1H, s), 7.34 (1H,s), 4.54 (2H, qd), 1.61 (9H, s), 1.46 (3H, t); m/z: 344+[M+H].

Intermediate 4

2-((2-Tert-butoxy-2-oxoethyl)(methyl)amino)-6-methoxyisonicotinic acid,shown below, was prepared as follows.

A slurry of 2-chloro-6-methoxyisonicotinic acid (100 mg, 0.53 mmol),sacrcosine tert-butyl ester (116 mg, (0.64 mmol),chloro(di-2-norborylphosphino)(2-dimethylaminomethylferrocen-1yl)palladium(II) (9.8 mg, 0.02 mmol) and sodium tert-butoxide (128 mg, 1.3 mmol) inp-dioxane (3 mL) were stirred at 110° C. for 18 hr. The reaction mixturewas cooled and the solvent removed in vacuo. The residue was dilutedwith water, pH adjusted to 4 with 1N aqueous hydrochloric acid andextrated with ethyl acetate (3×10 mL). The combined organic layer wasdried over magnesium sulfate, filtered, concentrated in vacuo and theresidue flash chromatographed (0-10% methanol/dichloromethane) to affordthe title compound (60 mg) as a yellow solid. m/z=297.5 (M+1).

Intermediate 5

2-(4-(Tert-butoxycarbonyl)phenyl)-6-methoxyisonicotinic acid, shownbelow, was prepared as follows.

2-Chloro-6-methoxyisonicotinic acid (15.0 g, 80.0 mmol), tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (29.2 g, 96.0mmol), 1,4-dioxane (500 mL) and sodium carbonate (25.4 g, 240 mmol)dissolved in water (160 mL) were combined in a 1 L, 3 necked flaskequipped with an internal thermometer, condenser and nitrogen inlet. Thesolution was degassed by bubbling with nitrogen for 15 min whilestirring. Tetrakis(triphenylphosphine)palladium (3.70 g, 3.20 mmol) wasthen added and the mixture heated to reflux for 17 h. The mixture wasthen cooled to room temperature and concentrated under vacuum to give athick brown suspension, which was portioned between ethyl acetate (400mL) and water (150 mL). The aqueous layer was separated and extractedwith further ethyl acetate (2×100 mL). The organic portions werecombined and washed with 1 N HCl and water, and the black solids removedby filtration through a pad of celite. The aqueous layer was discardedand the organic solution dried over anhydrous magnesium sulfate andfiltered. The solution was then treated with decolorlizing charcoal andheated to 70° C. for 20 min. The solution was then filtered throughcelite while hot and the solvent removed under vacuum to afford a yellowpowder. This material was purified by addition of methyl tert-butylether (55 mL) followed by the slow addition of 1.85 L heptane withstirring. The mixture was stirred for 2 days and then filtered to give2-(4-(tert-butoxycarbonyl)phenyl)-6-methoxyisonicotinic acid (22.01 g,84%) as a pale yellow powder. ¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 1.61(s, 9H) 4.08 (s, 3H) 7.34 (d, J=0.98 Hz, 1H) 7.97 (d, J=1.17 Hz, 1H)8.09 (s, 2H) 8.13 (s, 2H); m/z: 330.2+[M+H].

Intermediate 6

6-(4-(Tert-butoxycarbonyl)phenyl)-2-(methylamino)nicotinic acid, shownbelow, was prepared as follows.

Step 1. 6-Chloro-2-(methylamino)nicotinic acid, shown below, wasprepared as follows.

To a steel autoclave was added 2,6-dichloronicotinic acid (tech.) (30 g,156.2 mmoles), tetrahydrofuran (30 mL) and monomethylamine (68.2 mL. 33wt % in ethanol, 500 mmol). The reaction vessel was heated at 100° C.for 4 h. The reaction mixture was cooled and the solution transferredfrom the autoclave to a 500 mL flask and concentrated under vacuum togive a green solid. The solid was dissolved in 300 mL MeOH and 1.2 LEtOAc, poured into a separatory funnel and washed with 1N HCl (2×300 mL)and brine. The organic solution was then concentrated to dryness toyield 6-chloro-2-(methylamino)nicotinic acid (29 g, 96%) as an off-whitesolid.

1H NMR (400 MHz, DMSO-d6) δ ppm 13.14 (br. s., 1H), 8.17 (d, J=2.7 Hz,1H), 8.00 (d, J=8.0 Hz, 1H), 7.57 (br. s., 1H), 6.58 (d, J=8.0 Hz, 1H),2.89 (d, J=4.1 Hz, 3H).

m/z: 187+[M+H]; 185−[M−H]

Step 2. To a 2 L 3-neck flask was added6-chloro-2-(methylamino)nicotinic acid (22.9 g, 122.6 mmol), tert-butyl4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (31.2 g, 102.6mmol), 1,4-dioxane (1.02 L) and sodium carbonate (32.6 g, 307.7 mmol)dissolved in water (300 mL). The mixture was bubbled with dry nitrogenwhile stirring for 20 min. Tetrakis(triphenylphosphine)palladium (5.93g, 5.13 mmol) was then added and the mixture heated to reflux (89° C.)for 2 h. The reaction mixture was cooled to room temperature, water (250mL) added, and the mixture stirred for 10 min. The mixture was extractedwith 1.5 L of ethyl acetate and the organic layer separated and washedwith 10% aqueous sodium carbonate (250 mL), 1 N HCl (2×250 mL) andbrine. The solution was then concentrated to a minimum stirring volume,methanol (650 mL) added and the mixture heated at reflux to dissolve thesolids. 300 mL MeOH was removed by distillation and 100 mL water added.The mixture was then cooled to room temperature and the productcollected by filtration, washed with 150 mL of 2:1 methanol/water, anddried in the vacuum oven to afford the title compound (24 g, 71%) as ayellow solid. 1H NMR (400 MHz, CHLOROFORM-d) δ ppm 8.27 (d, J=8.2 Hz,1H), 8.13-8.21 (m, 2H), 8.03-8.13 (m, 2H), 7.82 (br. s., 1H), 7.11 (d,J=8.0 Hz, 1H), 3.21 (s, 3H), 1.63 (s, 9H); m/z: 329+[M+H]; 327−[M−H].

Intermediate 7

6-(3-(Tert-butoxycarbonyl)phenyl)-2-(ethylamino)nicotinic acid, shownbelow, was prepared as follows.

Step 1. 6-(3-(tert-butoxycarbonyl)phenyl)-2-chloronicotinic acid, shownbelow, was prepared as follows.

2,6-Dichloronicotinic acid (1.20 g, 6.25 mmol),3-(tert-butoxycarbonyl)benzene boronic acid (2.08 g, 9.10 mmol),potassium carbonate (2.60 g, 18.8 mmol),tetrakis(triphenylphosphine)palladium (0.72 g, 0.62 mmol), degassed1,2-dimethoxyethane (30 mL) and water (0.5 mL) were combined under anargon atmosphere and the mixture heated at 90° C. overnight. Thereaction mixture was then cooled to room temperature, diluted with water(100 mL) and extracted with ethyl acetate (2×150 mL). The aqueous layerwas acidified to pH 3-4 using 2N HCl solution and extracted with ethylacetate (2×200 mL). The combined organic portions were dried overanhydrous sodium sulfate and concentrated. The residue was purified bycolumn chromatography (silica gel, 20% methanol/dichloromethane) and theproduct containing fractions concentrated to give a solid, which wastriturated with 2:1 heptane/ethyl acetate to give pure6-(3-(tert-butoxycarbonyl)phenyl)-2-chloronicotinic acid (1.4 g, 67%) asan off-white foam. ¹H NMR (400 MHz, CD₃OD) δ ppm 8.64 (s, 1H), 8.34 (d,1H), 8.29 (d, 1H), 8.04 (d, 1H), 7.95 (d, 1H), 7.59 (t, 1H), 1.60 (s,9H); m/z 334 [M+H].

Step 2. 6-(3-(Tert-butoxycarbonyl)phenyl)-2-chloronicotinic acid (500mg, 1.50 mmol) was dissolved in a 2 M solution of ethylamine intetrahydrofuran (7.0 mL, 14.0 mmol) and the mixture heated using amicrowave at 140° C. for 4 h. The solution was then concentrated todryness and the crude mixture purified by column chromatography (silicagel, 10% methanol/dichloromethane). The solid obtained was trituratedwith ethyl acetate/heptane (1:4) and filtered to afford the titlecompound (330 mg, 64% yield) as a yellow solid. ¹H NMR (400 MHz, CDCl₃)δ ppm 8.70 (s, 1H), 8.27 (dd, 2H), 8.06 (d, 1H), 7.85 (br.s, 1H), 7.52(t, 1H), 7.09 (d, 1H), 3.71 (q, 1H), 1.62 (s, 9H), 1.27 (t, 3H); m/z 343[M+H].

Intermediate 8

4-(3-(Tert-butoxycarbonyl)phenyl)-1H-pyrrolo[2,3-b]pyridine-2-carboxylicacid, shown below, was prepared as follows:

Step 1. Methyl4-(3-(tert-butoxycarbonyl)phenyl)-1H-pyrrolo[2,3-b]pyridine-2-carboxylate,shown below, was prepared as follows:

A stirred slurry of methyl 4-chloro-7-azaindole-2-carboxylate (100 mg,0.48 mmol), tert-butyl 3-(hydroxy(methyl)boryl)benzoate (137 mg, 0.62mmol), potassium phosphate (309 mg, 1.42 mmol),tetrakis(triphenylphosphine) palladium (28 mg, 0.02 mmol) in p-dioxane(3 mL)/water (1 mL) were heated at 100° C. for 15 hr. The reactionmixture was cooled, concentrated in vacuo and flash chromatographed(0-100% ethyl acetate:heptanes) to afford methyl4-(3-(tert-butoxycarbonyl)phenyl)-1H-pyrrolo[2,3-b]pyridine-2-carboxylateas a white powder (148 mg). 1H NMR (400 MHz, CHLOROFORM-d) d ppm 1.62(s, 9H) 3.99 (s, 3H) 7.28 (d, J=5.23 Hz, 1H) 7.36 (d, J=2.62 Hz, 1H)7.59 (t, 1H) 7.90 (d, 1H) 8.09 (d, 1H) 8.35 (t, 1H) 8.64 (d, J=4.70 Hz,1H); m/z (M+1)=353.2.

Step 2. To a stirred solution of methyl4-(3-(tert-butoxycarbonyl)phenyl)-1H-pyrrolo[2,3-b]pyridine-2-carboxylate(145 mg, 0.41 mmol) in methanol (2 mL)/tetrahydrofuran (2 mL) was added1N aqueous lithium hydroxide (0.82 mL, 0.82 mmol). After 18 hr, thereaction mixture was concentrated in vacuo, diluted into water, pHadjusted to ˜5 with 1N aqueous hydrochloric acid, the yellow solidscollected by filtration and dried in vacuo to afford the title compound(122 mg). m/z=339.5 (M+1); ¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.56 (s, 1H)8.47 (d, J=4.88 Hz, 1H) 8.24 (t, J=1.66 Hz, 1H) 8.01 (tt, J=8.36, 1.29Hz, 2H) 7.69 (t, J=7.80 Hz, 1H) 7.31 (d, J=4.88 Hz, 1H) 7.15 (d, J=2.15Hz, 1H) 1.52-1.59 (m, 9H).

Intermediate 9

1-isopropyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one, shownbelow, was prepared as follows.

Step 1. tert-butyl 9-oxo-3-azaspiro[5.5]undec-7-ene-3-carboxylate, shownbelow, was prepared as follows.

Methyl vinyl ketone (146 mL) was added to a solution of tert-butyl4-formylpiperidine-1-carboxylate (375 g) in tetrahydrofuran (18 L). Thereaction mixture was cooled to −5° C. and a solution of potassiumhydroxide in ethanol (3N, 0.243 L) was added dropwise over 10 minutes.The reaction mixture was allowed to warm to room temperature and stirredfor 16 hours. Cyclohexane (10 L) was added and the solution was washedwith saturated sodium chloride (3×10 L). The organic layer wasconcentrated to an oil. This oil was dissolved in 2 L of 80:20cyclohexane/ethyl acetate and filtered through Celite® to removeinsoluble material. The filtrate was purified via flash columnchromatography (70:30 hexane/ethyl acetate) to afford the product as anoil. The oil was triturated in hexanes to afford the desired product asa white solid (131 g, 28%).

Step 2. (E)-tert-butyl10-((dimethylamino)methylene)-9-oxo-3-azaspiro[5.5]undec-7-ene-3-carboxylate,shown below, was prepared as follows.

tert-Butyl 9-oxo-3-azaspiro[5.5]undec-7-ene-3-carboxylate (101 g) andtris(dimethylaminomethane) (133 mL) were dissolved in toluene (800 mL)and heated to reflux for 17 hours. The reaction mixture was concentratedto a minimum stirring volume and ethyl acetate (600 mL) was added. Thismixture was heated to reflux and heptane (1.2 L) was added over 20minutes. The hot solution was cooled to room temperature over 3 hours.The solids were filtered through a course glass frit and washed withheptane (300 mL). The resulting solid was dried in a vacuum oven at 40°C. for 3 hours to afford the desired product as a yellow solid (107 g).¹H NMR (400 MHz, CDCl₃) δ ppm 7.48 (s, 1H), 6.57 (d, J=9.97 Hz, 1H),5.99 (d, J=10.16 Hz, 1H), 3.32-3.51 (m, 4H), 3.06 (s, 6H), 2.72 (s, 2H),1.57-1.66 (m, 2H), 1.41-1.53 (m, 11H).

Step 3. tert-Butyl1-isopropyl-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

(E)-tert-Butyl10-((dimethylamino)methylene)-9-oxo-3-azaspiro[5.5]undec-7-ene-3-carboxylate(107 g) was taken up in toluene (700 mL) and isopropyl hydrazine (44.4g) was added. The reaction was stirred at reflux for 4 hours. Thereaction was cooled to room temperature and ethyl acetate was added (500mL). The reaction solution was washed with citric acid (2×300 mL, 10%aqueous), and water (400 mL). The organic layer concentrated in vacuo toafford tert-butyl1-isopropyl-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate asa yellow solid (109 g). ¹H NMR (400 MHz, CDCl₃) δ ppm 7.25 (s, 1H) 6.42(dd, J=10.05, 0.49 Hz, 1H) 5.84 (d, J=9.95 Hz, 1H) 4.42-4.52 (m, 1H)3.36-3.53 (m, 4H) 2.62 (s, 2H) 1.56-1.68 (m, 2H) 1.45-1.55 (m, 17H).

Step 4. tert-butyl1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

To a solution of tert-butyl1-isopropyl-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(109 g) in methanol (1 L) was added N-bromo succinimide (61.4 g). Thereaction was stirred for 1 hour. The reaction was quenched with sodiumthiosulfate (10 g in 300 mL water) and then distilled to a final volumeof 500 mL. The solution was cooled to ambient temperature and 2-methyltetrahydrofuran (1 L) and water (100 mL) were added. The organic layerwas removed and the aqueous layer was extracted with 2-methyltetrahydrofuran. The organic layers were combined, washed with aqueoussodium hydroxide (1 N, 250 mL), water and saturated, aqueous sodiumchloride. The organic layer was dried over sodium sulfate, filtered andconcentrated to an orange oil. The oil was dissolved in tetrahydrofuran(500 mL) and potassium tert-butoxide (76.8 g) in tetrahydrofuran (500mL) was added. The solution was heated to 60° C. and stirred for 1 hour.Aqueous hydrochloric acid (1 N, 1 L) was added and the solution wasstirred for 30 minutes. The phases were separated and the aqueous layerwas extracted with ethyl acetate (700 mL). The organic layers werecombined, washed with water (400 mL) and saturated, aqueous sodiumchloride (100 mL). The organic layer was dried over sodium sulfate,filtered and concentrated in vacuo to give a residue. The residue wasdried in a vacuum oven at 40° C. for 16 hours to afford the titlecompound as an orange wax (117 g). ¹H NMR (400 MHz, CDCl₃) δ ppm 7.35(s, 1H), 5.32-5.42 (m, 1H), 3.29-3.51 (m, 4H), 2.73 (s, 2H), 2.51 (s,2H), 1.47-1.57 (m, 4H), 1.42-1.46 (m, 15H); +ESI MS (M+H)=348.5.

Step 5. 1-isopropyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one,shown below, was prepared as follows.

tert-Butyl 9-oxo-3-azaspiro[5.5]undec-7-ene-3-carboxylate (250 g), andtris(dimethylaminomethane) (325 mL) were dissolved in toluene (1.9 L)and heated at reflux for 4 hours. The mixture was distilled andconcentrated to a minimum stirring volume (110° C.) and then toluene(1.9 L) was added. The reaction was redistilled to a minimum stirringvolume and cooled to room temperature. Toluene (1.8 L) and isopropylhydrazine hydrochloride (135 g) were added and the solution was heatedto reflux for 5 hours. The reaction was cooled to room temperature andwas then washed with citric acid (10% aqueous, 2×150 mL) and water (200mL), and then the organic layer was distilled to a minimum stirringvolume. Methanol (2 L) was added and distilled to a minimum stirringvolume. This was repeated with methanol (2 L). The solution wasredissolved in methanol (2.5 L) and N-bromosuccimimide (176 g) was addedin one portion. The solution was stirred at 23° C. for 2 hours. Aqueoussodium thiosulfate solution (5 wt %, 0.5 L) was added and the mixturewas stirred for 15 minutes. The reaction mixture was concentrated viadistillation (45° C., 210 mm Hg) to ˜0.5 L and then 2-methyltetrahydrofuran (2.5 L) was added. After stirring for 15 minutes theaqueous layer was discarded. The organic layer was concentrated to ˜0.2L and tetrahydrofuran (0.5 L) was added. To the mixture was added apotassium tert-butoxide solution in tetrahydrofuran (1.9 L, 1 Msolution). The solution was heated to 60° C. and stirred for 1 hour.After cooling to room temperature, aqueous hydrochloric acid (1 N, 2.2L) was added over 20 minutes. The mixture was stirred at roomtemperature for 20 minutes, and then the layers were allowed toseparate. The aqueous layer was removed and back extracted with ethylacetate (1.75 L). The combined organic layers were washed with water (1L) and the organic layer concentrated via distillation (4 L solventremoved). Ethyl acetate (1.8 L) was added and the solution wasconcentrated to a minimum stirring volume. Ethyl acetate (3 L) andmethanol (0.8 L) were added and the solution was cooled to 0° C. Acetylchloride (401 mL) was added dropwise over 20 minutes and the solutionwas stirred at 0° C. for 4 hours. The precipitate was collected byfiltration under nitrogen. The filtrate was washed with ethyl acetate(0.5 L) and dried in a vacuum oven at 40° C. to afford1-isopropyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one as anoff-white solid (241 g). ¹H NMR (400 MHz, CD₃OD) δ ppm 7.43 (s, 1H),5.32-5.42 (m, 1H), 3.15-3.25 (m, 4H), 2.89 (s, 2H), 2.64 (s, 2H),1.69-1.90 (m, 4H), 1.37-1.45 (m, 6H); +ESI (M+H)=248.4

Intermediate 10

1-tert-butyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one, shownbelow, was prepared as follows.

Step 1. Benzyl10-((dimethylamino)methylene)-9-oxo-3-azaspiro[5.5]undec-7-ene-3-carboxylate,shown below, was prepared as follows.

9-oxo-3-aza-spiro[5.5]undec-7-ene-3-carboxylic acid benzyl ester (15.2g, 51 mmol) was dissolved in 180 mL toluene andtris(dimethylamino)methane (22.2 g, 27 mmol) was added. The reaction washeated to reflux for 5 hours and then allowed to cool to roomtemperature overnight. The reaction solution was concentrated in vacuoto provide the title compound (18.0 g, 100%): +APCI MS (M+H) 354.6; ¹HNMR (400 MHz, CDCl₃) δ ppm 7.49 (s, 1H), 7.28-7.40 (m, 5H), 6.59 (d,J=10.16 Hz, 1H), 6.01 (d, J=9.97 Hz, 1H), 5.13 (s, 2H), 3.52-3.66 (m,2H), 3.39-3.52 (m, 2H), 3.07 (s, 6H), 2.74 (s, 2H), 1.58-1.73 (m, 2H),1.41-1.58 (m, 2H).

Step 2. Benzyl1-tert-butyl-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl10-((dimethylamino)methylene)-9-oxo-3-azaspiro[5.5]undec-7-ene-3-carboxylate(59.2 g, 167 mmol) was dissolved in 835 mL of ethanol. To the reactionsolution was added acetic acid (20 mL, 345 mmol) and tert-butylhydrazinehydrochloride (29.1 g, 234 mmol). The reaction was heated to reflux for1 hour. The reaction solution was cooled to room temperature and thenconcentrated in vacuo to give an orange oil which was purified by flashchromatography using 20-40% ethyl acetate in heptane as eluent to affordthe title compound as a pale yellow solid (50 g, 79%): +ESI MS (M+H)380.5; ¹H NMR (400 MHz, CDCl₃) δ ppm 7.26-7.40 (m, 5H) 7.17 (s, 1H) 6.66(d, J=9.95 Hz, 1H) 5.77 (d, J=10.15 Hz, 1H) 5.12 (s, 2H) 3.38-3.64 (m,4H) 2.58 (s, 2H) 1.60 (s, 12H) 1.50 (br. s., 1H).

Step 3. Benzyl6-bromo-1-tert-butyl-7-hydroxy-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl1-tert-butyl-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(50 g, 132 mmol) was dissolved in 1 L of tetrahydrofuran. To thereaction was added N-bromosuccinimide (24.6 g, 138 mmol) and 250 mL ofwater. The reaction was stirred for 1 hour at room temperature. Thereaction was partitioned between ethyl acetate and water. The phaseswere separated and the organic phase was washed an additional 2 timeswith water and once with saturated, aqueous sodium chloride. The organicphase was dried over magnesium sulfate, concentrated in vacuo, andcrystallized from ether to afford the title compound as a cream-coloredsolid (60.7 g, 97%): +ESI MS (M+H) 476.5; ¹H NMR (400 MHz, CDCl₃) δ ppm7.28-7.36 (m, 5H), 7.27 (s, 1H), 5.23 (t, J=4.68 Hz, 1H), 5.12 (s, 2H),4.24 (d, J=4.49 Hz, 1H), 3.87 (br. s., 2H), 3.12 (br. s., 2H), 2.79 (d,J=16.00 Hz, 2H), 2.59 (d, J=15.80 Hz, 2H), 1.95 (br. s., 1H), 1.66 (s,11H), 1.58 (br. s., 1H).

Step 4. Benzyl6-bromo-1-tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl6-bromo-1-tert-butyl-7-hydroxy-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(57.9 g, 122 mmol) was dissolved in 1 L acetone and then cooled to 0° C.in an ice bath. To the solution was added 122 mL of Jones Reagent(Fillion, E. Tetrahedron Letters 2004, 46, 1091-1094). The ice bath wasremoved and the reaction was allowed to warm to room temperature whereit was stirred for 45 minutes. Saturated, aqueous sodium bicarbonate wasadded until no further gas evolution was noted and pH reached 7. Theresulting mixture was filtered through a pad of Celite® rinsing withethyl acetate. The filtrate layers were separated and the aqueous layerwas back extracted with ethyl acetate. The organic extracts werecombined, washed twice with water, once with saturated aqueous sodiumchloride, dried over magnesium sulfate and concentrated in vacuo. Theresidue was crystallized from ethyl acetate/heptane to afford the titlecompound (50.4 g, 87%): +ESI MS (M+H) 474.5; ¹H NMR (400 MHz, CDCl₃) δppm 7.32 (d, J=9.38 Hz, 6H), 5.11 (s, 2H), 4.24 (s, 1H), 3.58-3.84 (m,2H), 3.16-3.41 (m, 2H), 2.67-2.91 (m, 2H), 1.80 (br. s., 1H), 1.61-1.76(m, 11H), 1.52-1.61 (m, 1H).

Step 5. Benzyl1-tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl6-bromo-1-tert-butyl-7-hydroxy-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(50.4 g, 106 mmol) was dissolved in 600 mL of tetrahydrofuran, to thiswas added saturated, aqueous ammonium chloride (600 mL) and zinc powder(20.8 g, 319 mmol). The reaction was stirred for 30 minutes at roomtemperature. The reaction was filtered through Celite®, the phases wereseparated and the organic phase was washed with water and saturated,aqueous sodium chloride, dried over magnesium sulfate and concentratedin vacuo to give a foam. The foam was triturated once in ethylacetate/heptane and once in ether and filtered to afford the titlecompound as a white solid (40.4 g, 96%): +ESI MS (M+H) 396.5; ¹H NMR(400 MHz, CDCl₃) δ ppm 7.24-7.38 (m, 6H), 5.11 (s, 2H), 3.36-3.61 (m,4H), 2.74 (s, 2H), 2.54 (s, 2H), 1.64 (s, 9H), 1.51 (br. s., 4H).

Step 6.1-tert-butyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one, shownbelow, was prepared as follows.

Benzyl1-tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(46.6 g, 118 mmol) was dissolved in 730 mL ethanol and the solution wasadded to 10% palladium on carbon (9.4 g). To this was added1-methyl-1,4-cyclohexadiene (90 mL, 769 mmol). The reaction was stirredat reflux for 2 hours. The reaction was cooled to room temperature andfiltered through Celite®. The filtrate was concentrated in vacuo to givea gray solid. The solid was dissolved in 150 mL ethyl acetate, to thiswas added 35 mL 4 M hydrochloric acid in dioxane. A precipitate formedand was collected by filtration to afford the title compound as a whitesolid (34 g, 97%): +ESI MS (M+H) 262.5; ¹H NMR (400 MHz, CD₃OD) δ ppm7.34 (s, 1H) 3.12-3.25 (m, 4H) 2.90 (s, 2H) 2.66 (s, 2H) 1.67-1.85 (m,4H) 1.62 (s, 9H).

Intermediate 11

1-(Oxetan-3-yl)-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one,shown below, was prepared as follows.

Step 1. tert-Butyl1-(2-ethoxy-2-oxoethyl)-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Ethylhydrazinoacetate hydrochloride (0.92 g, 5.95 mmol) was added to asolution of benzyl10-((dimethylamino)methylene)-9-oxo-3-azaspiro[5.5]undec-7-ene-3-carboxylate(1.25 g, 3.90 mmol), described in the preparation of Intermediate 10, inethanol (30 mL). Stir the mixture at reflux for 1 hour. An aliquotindicated the reaction was complete by ¹HNMR. The reaction mixture wascooled to room temperature and concentrated under high vacuum to a brownoil. The oil was triturated with diethyl ether (50 mL). The precipitatewas filtered and the filtrate concentrated and dried under high vacuumto yield the title compound (1.50 g, 100%) as a brown oil. +APCI MS(M+H) 376.2; ¹H NMR (400 MHz, CDCl₃) δ ppm 1.21-1.26 (m, 3H), 1.43 (s,9H), 1.45-1.52 (m, 2H), 1.54-1.64 (m, 2H), 2.62 (s, 2H), 3.33-3.49 (m,4H), 4.15-4.22 (m, 2H), 4.82 (s, 2H), 5.87 (d, J=9.97 Hz, 1H), 6.26 (d,J=9.97 Hz, 1H), 7.24 (s, 1H).

Step 2. Diethyl2-(1′-(tert-butoxycarbonyl)spiro[indazole-5,4′-piperidine]-1(4H)-yl)malonate,shown below, was prepared as follows.

tert-Butyl1-(2-ethoxy-2-oxoethyl)-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(1.45 g 3.86 mmol) in toluene (5 mL) was added to a suspension of sodiumhydride (0.148 g, 60% dispersion in mineral oil) in diethyl carbonate(30 mL), dropwise at 80° C. over 30 minutes. The reaction was stirred atreflux for 1.5 hours. ¹H NMR indicated that starting material wasconsumed and that the desired product had formed. The reaction mixturewas cooled to room temperature. Methanol (1 mL) was added and thesolution was stirred at room temperature for 5 minutes. Water (5 mL) wasadded. The solution was acidified to pH˜3 with 2 N aqueous, hydrochloricacid (3 mL) then was extracted with dichloromethane (3×15 mL). Thecombined organics were dried over magnesium sulfate, filtered,concentrated, and dried under high vacuum to yield a brown gum (1.59 g,92%). The crude material was triturated with 1:1 diethyl ether:heptanes(50 mL). The precipitate was filtered. The filtrate was concentrated anddried under high vacuum to yield the title compound (1.25 g, 72%). +APCIMS (M+H) 348.1; ¹H NMR (400 MHz, CDCl₃) δ ppm 1.13-1.32 (m, 6H),1.40-1.46 (m, 9H), 1.46-1.54 (m, 2H), 1.59 (d, J=13.68 Hz, 3H), 2.62 (s,2H), 3.31-3.51 (m, 4H), 4.27 (q, J=7.23 Hz, 4H), 5.85 (d, J=9.97 Hz,1H), 6.34 (d, J=9.97 Hz, 1H), 7.24 (s, 1H).

Step 3. tert-Butyl1-(1,3-dihydroxypropan-2-yl)-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Tetrahydrofuran (40 mL) was added to lithium aluminum hydride (900 mg)in a 3-neck, 125 mL roundbottom flask equipped with a nitrogen inlet andthermometer. The solution was cooled to −2° C. Diethyl2-(1′-(tert-butoxycarbonyl)spiro[indazole-5,4′-piperidine]-1(4H)-yl)malonate(1 g) in tetrahydrofuran (5 mL) was added dropwise over 5 minutes. Thetemperature was never greater than −0.2° C. during the addition. Thereaction was stirred at 0° C. for 3 hours then the reaction was thenquenched through the sequential addition of water (1.0 mL), 15% aqueoussodium hydroxide (1.0 mL), and water (3 mL). The internal temperaturewas never greater than 3.2° C. during the addition. The reaction wasthen allowed to warm to room temperature over 15 minutes. The reactionmixture was filtered through Celite® and washed with diethyl ether (3×20mL). The combined organics were washed with brine (5 mL) dried oversodium sulfate, filtered, concentrated, and dried under high vacuum toyield a pale yellow glass (548 mg, 67%). This material waschromatographed on 25 g of silica eluting with 2% to 8% methanol indichloromethane with 0.1% ammonium hydroxide over 30 minutes to yieldthe title compound (133 mg, 16%). +APCI MS (M+H) 364.2; ¹H NMR (400 MHz,CDCl₃) δ ppm 1.45 (s, 9H), 1.51 (br. s., 2H), 1.60 (br. s., 4H), 2.62(s, 2H), 3.32-3.53 (m, 4H), 4.05 (br. s., 4H), 4.26 (t, J=4.89 Hz, 1H),5.89 (s, 1H), 6.40 (d, J=9.77 Hz, 1H), 7.23-7.25 (m, 1H).

Step 4. tert-Butyl1-(oxetan-3-yl)-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

2.5 M n-butyl lithium in hexanes (0.33 ml165 uL) was added to a solutionof tert-Butyl1-(1,3-dihydroxypropan-2-yl)-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(150 mg, 0.41 mmol) in tetrahydrofuran (8 mL) at −6.2° C. Thetemperature was never greater than −3.7° C. during the addition. Thesolution was stirred at −8° C. for 30 minutes. A solution ofp-toluenesulfonyl chloride (79 mg) in tetrahydrofuran (2 mL) was addedto the reaction mixture at −5° C. The temperature was never greater than−2° C. during the addition. The reaction was stirred at −5° C. for 1hour then the reaction mixture was cooled to −6° C. and 2.5 M n-butyllithium in hexanes (0.33 mL, 165 uL) was added over 2 minutes. Thetemperature was never greater than −3.5° C. during the addition. Thecooling bath was removed and the reaction was stirred at an internaltemperature of 60° C. for 16 hours. The reaction mixture was cooled toroom temperature and ethyl acetate (20 mL) was added. The reactionsolution was washed with water (35 mL) and the aqueous layer wasextracted with ethyl acetate (15 mL). The combined organics were washedwith brine (5 mL) dried over magnesium sulfate, filtered, concentrated,and dried under high vacuum to yield a yellow solid. The solid waspurified by chromatography on 8 g silica eluting with 25% to 75% ethylacetate in heptanes over 36 minutes to yield the title compound (58 mg,40%). +ESI MS (M+H); ¹H NMR (400 MHz, CDCl₃) δ ppm 1.45 (s, 9H), 1.49(d, J=3.71 Hz, 1H), 1.55 (s, 4H), 1.59 (br. s., 1H), 2.61 (s, 2H),3.32-3.50 (m, 4H), 5.00 (m, J=7.22, 7.22 Hz, 2H), 5.13 (t, J=6.44 Hz,2H), 5.36-5.46 (m, 1H), 5.88 (d, J=9.95 Hz, 1H), 6.43 (d, J=9.95 Hz,1H), 7.33 (s, 1H).

Step 5. tert-Butyl6-bromo-7-methoxy-1-(oxetan-3-yl)-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

N-Bromosuccinimide (30 mg, 0.17 mmol) was added to tert-butyl1-(oxetan-3-yl)-1,4-dihydrospiro[indazole-5,4′-piperidine]-t-carboxylate(56 mg, 0.17 mmol) in methanol (1.0 mL) at room temperature. Thereaction was stirred at room temperature for 2 hours thenN-bromosuccinimide (4.5 mg) was added and the reaction mixture wasstirred at room temperature for 1 hour. The reaction mixture wasconcentrated under a stream of nitrogen to a residue. Ethyl acetate (15mL) was added and the reaction solution was washed with 10% citric acid(3 mL), 1N sodium hydroxide (3 mL), and brine (3 mL). The organic layerwas concentrated and dried under high vacuum to yield the title compound(74 mg, 100%) as a colorless solid. +APCI MS (M+H) 458.2; ¹H NMR (400MHz, CDCl₃) δ ppm 1.44 (s, 9H), 1.69 (br. s., 4H), 2.51 (d, J=15.83 Hz,1H), 2.67 (d, J=15.83 Hz, 1H), 3.06-3.31 (m, 3H), 3.54 (s, 3H),3.62-3.72 (m, 1H), 4.39 (s, 1H), 4.66 (s, 1H), 4.87-4.93 (m, 1H), 4.97(t, J=6.84 Hz, 1H), 4.99-5.04 (m, 1H), 5.30 (s, 1H), 5.34-5.40 (m, 1H),7.43 (s, 1H).

Step 6. tert-Butyl1-(oxetan-3-yl)-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

1 M potassium tert-butoxide in tetrahydrofuran (0.320 mL) was added to asolution of tert-butyl6-bromo-7-methoxy-1-(oxetan-3-yl)-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate72 mg, 0.16 mmol) in tetrahydrofuran (1.0 mL) at room temperature. Thecolorless solution turned yellow upon addition. The solution was stirredat room temperature for 16 hours. 1 N aqueous, hydrogen chloride (0.475mL, 3 eq.) was added and the solution was stirred at room temperaturefor 1 hour. The tetrahydrofuran was concentrated under a stream ofnitrogen. The aqueous phase was extracted with ethyl acetate (3×5 mL).The combined organics were washed with brine (3 mL) then the organiclayer was concentrated and dried under high vacuum to give the titlecompound as a pale yellow solid (54 mg, 96%). −APCI MS (M−H) 360.2; ¹HNMR (400 MHz, CD₃OD) δ ppm 1.38-1.45 (m, 9H), 1.46-1.56 (m, 4H), 2.57(s, 2H), 2.82 (s, 2H), 3.33-3.53 (m, 4H), 4.94-5.06 (m, 4H), 6.08-6.21(m, 1H), 7.53 (s, 1H).

Step 7.1-(oxetan-3-yl)-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one,shown below, was prepared as follows.

Trifluoroacetic acid (0.2 mL) was added to a solution of tert-butyl1-(oxetan-3-yl)-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(50 mg, 0.14 mmol) in dichloromethane (2 mL) at 0° C. The cooling bathwas removed and the reaction was stirred at room temperature for 1.5hours. The reaction mixture was concentrated to a residue under a streamof nitrogen and dried under high vacuum for 20 minutes. The residue wastriturated with diethyl ether (5 mL). The solvent was decanted and theresulting precipitate was dried under high vacuum to yield the titlecompound (52 mg, 100) as a pale yellow solid. +APCI MS (M+H) 262.2; ¹HNMR (400 MHz, CD₃OD) δ ppm 1.65-1.86 (m, 4H), 2.63 (s, 2H), 2.89 (s,2H), 3.14-3.27 (m, 4H), 5.02 (s, 4H), 6.07-6.21 (m, 1H), 7.53-7.60 (m,1H).

Intermediate 12

1-Isopropyl-3-methyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one,shown below, was prepared as follows.

Step 1. Benzyl1-isopropyl-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl10-((dimethylamino)methylene)-9-oxo-3-azaspiro[5.5]undec-7-ene-3-carboxylate(6.38 g, 18 mmol), prepared as described in the preparation ofIntermediate 10, was dissolved in 90 mL of ethanol. To the reactionsolution was added acetic acid (2.16 g, 36 mmol) and1-isopropylhydrazine hydrochloride (2.79 g, 25 mmol). The reaction washeated to reflux for 2 hours then the reaction solution was cooled toroom temperature and concentrated in vacuo to give an orange oil whichwas purified by flash chromatography using 12-100% ethyl acetate inheptane as eluent to afford the title compound as a yellow gum (6.58 g,69%): +ESI MS (M+H) 366.5; ¹H NMR (400 MHz, CDCl₃) δ ppm 7.28-7.39 (m,5H), 7.25 (s, 1H), 6.42 (d, J=9.95 Hz, 1H), 5.84 (d, J=9.95 Hz, 1H),5.14 (s, 2H), 4.41-4.54 (m, 1H), 3.42-3.65 (m, 4H), 2.62 (s, 2H),1.58-1.70 (m, 2H), 1.50-1.58 (m, 2H), 1.49 (d, J=6.83 Hz, 6H).

Step 2. Benzyl3,6-dibromo-1-isopropyl-7-methoxy-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl1-isopropyl-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(679 mg, 1.86 mmol) was dissolved in 15 mL methanol and treated withN-bromosuccinimide (728 mg, 4.09 mmol) and the reaction was stirred atambient temperature for 18 hours. The methanol was removed under reducedpressure. The resultant tan foam was taken up in 50 mL ethyl acetate andwashed with 0.5 M sodium hydroxide (2×50 mL) and 20 mL saturated aqueoussodium thiosulfate. The organic phase was dried over sodium sulfate,filtered and concentrated. The resultant oil was flash chromatographed(25 g silica, 10-80% ethyl acetate/heptane gradient) to yield 784 mg(76%) of the title compound as a white foam: +APCI−MS (M+H)=554.1,556.2, 558.2: ¹H NMR (400 MHz, CDCl₃) δ ppm 7.26-7.42 (m, 5H), 5.12 (s,2H), 4.67 (d, J=1.76 Hz, 1H), 4.36 (s, 1H), 4.27 (m, 1H), 3.79 (d,J=11.90 Hz, 1H), 3.59-3.73 (m, 1H), 3.53 (s, 3H), 3.24-3.40 (m, 1H),3.19 (ddd, J=13.61, 10.00, 3.12 Hz, 1H), 2.56 (d, J=16.19 Hz, 1H), 2.34(d, J=16.19 Hz, 1H), 1.56-1.85 (m, 4H), 1.38-1.55 (m, 6H).

Step 3. Benzyl3-bromo-1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl3,6-dibromo-1-isopropyl-7-methoxy-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(784 mg, 1.4 mmol) was dissolved in 15 mL tetrahydrofuran. Potassiumt-butoxide (2.82 mL, 2 eq, 1 M tetrahydrofuran) was added and thereaction was stirred for 18 hours at ambient temperature. To thereaction was added 25 mL 2 N hydrochloric acid. The mixture was stirredfor 30 minutes at ambient temperature. The mixture was diluted with 25mL water and extracted with ethyl acetate (2×50 mL). The organicextracts were combined and dried over sodium sulfate, filtered andconcentrated. The resultant oil was flash chromatographed (50 g silica,8-66% ethyl acetate/heptane gradient) to yield 612 mg of the titlecompound as a white foam: +ESI MS (M+H)=462.5 ¹H NMR (400 MHz, CDCl₃) δppm 7.25-7.38 (m, 5H), 5.24-5.42 (m, 1H), 5.12 (s, 2H), 3.49-3.66 (m,2H), 3.46 (dd, J=7.41, 4.88 Hz, 2H), 2.63 (s, 2H), 2.52 (s, 2H),1.48-1.65 (m, 4H), 1.44 (d, J=6.63 Hz, 6H).

Step 4. tert-Butyl3-bromo-1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl3-bromo-1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(612 mg, 1.33 mmol) was dissolved in 10 mL 33% hydrobromic acid/aceticacid and the mixture was stirred for 60 minutes at ambient temperature.The solvent was evaporated and the red-orange residue taken up in 50 mLwater and made basic with saturated aqueous sodium carbonate andextracted with ethyl acetate (2×50 mL). The organic phase wasconcentrated to 20 mL and treated with 20 mL saturated aqueous sodiumbicarbonate and di-tert-butyl dicarbonate (348 mg). The biphasic mixturestirred for one hour at ambient temperature. The layers were separatedand the organic phase dried over sodium sulfate, filtered andconcentrated under reduced pressure. The resultant oil was flashchromatographed (10-70% ethyl acetate/heptane, 10 g silica) to yield 364mg of the title compound. +ESI MS (M+H)=413.5; ¹H NMR (400 MHz, CDCl₃) δppm 5.24-5.43 (m, 1H), 3.41-3.56 (m, 2H), 3.28-3.41 (m, 2H), 2.63 (s,2H), 2.51 (s, 2H), 1.47-1.56 (m, 4H), 1.40-1.49 (m, 15H).

Step 5. tert-Butyl1-isopropyl-3-methyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

tert-Butyl3-bromo-1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(440 mg, 1.03 mmol), palladium tetrakis triphenylphosphine (119 mg,0.103 mmol), potassium carbonate (146 mg, 1.03 mmol), and water (94 mg,5.16 mmol) were combined in dimethylformamide (2 mL) and degassed withnitrogen for 2 minutes. The reaction vial was sealed and heated in amicrowave reactor for 30 minutes at 100° C. The vial was removed fromthe microwave reactor and then heated to 100° C. in a conventionalheating block for 4 days. The reaction was concentrated in vacuo andthen partitioned between water (5 mL) and ethyl acetate (5 mL). Thephases were separated and the organic layer was concentrated and thenchromatographed on a 40 g column eluting with 20-40% ethyl acetate inheptane gradient to give 268 mg (72%) of the title compound. +ESI MS(M+H)=362.5; ¹H NMR (400 MHz, CDCl₃) δ ppm 5.20-5.53 (m, 1H), 3.32-3.54(m, 4H), 2.62 (s, 2H), 2.50 (s, 2H), 2.23 (s, 3H), 1.53 (t, J=5.76 Hz,4H), 1.46 (s, 9H), 1.44 (d, J=6.64 Hz, 6H).

Step 6.1-Isopropyl-3-methyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one,shown below, was prepared as follows.

tert-Butyl1-isopropyl-3-methyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(375 mg, 1.04 mmol) was dissolved in 3 mL diethyl ether and treated with4 M hydrogen chloride in dioxane (1 mL). The solution was stirred forone hour and then concentrated in vacuo to provide 300 mg of the titlecompound as a white foam: ¹H NMR (400 MHz, DMSO-d₆) δ ppm 5.10-5.35 (m,1H), 4.34 (br. s., 4H), 2.70 (s, 2H), 2.56 (s, 2H), 2.17 (s, 3H), 1.66(br. s., 4H), 1.34 (d, J=6.64 Hz, 6H).

Intermediate 13

1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-3-carbonitrile,shown below, was prepared as follows.

Step 1. tert-Butyl3-cyano-1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

In a schlenk tube flushed with nitrogen was added tert-butyl3-bromo-1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(250 mg, 0.59 mmol), prepared as described in the preparation ofIntermediate 12, tris(dibenzylideneacetone)dipalladium(0)-chloroformadduct (23.8 mg, 0.02 mmol), zinc dust (9.6 mg, 0.15 mmol), zinc cyanide(75.7 mg, 0.65 mmol) and 1,1′-bis(diphenylphosphino)ferrocene (26.1 mg,0.05 mmol). Anhydrous dimethylacetamide (3.5 mL) was added and the flaskwas flushed with nitrogen, then capped with a Teflon® screw top. Thereaction was stirred at 120° C. for 16 hours. The reaction was cooledand then filtered through a pad of Celite® washing with ethyl acetate.The filtrate was washed with water and the aqueous phase was backextracted with ethyl acetate. The combined organic phases were washedwith saturated aqueous sodium chloride, dried over magnesium sulfate andconcentrated in vacuo. The residue was purified by flash chromatographyusing 5-30% ethyl acetate in heptane gradient to give 204 mg of thetitle compound as a solid (93%): +ESI MS (M-Boc+H) 273.5; ¹H NMR (400MHz, CD₃OD) δ ppm 5.44 (m, 1H), 3.44 (m, 4H), 2.89 (s, 2H), 2.64 (s,2H), 1.53 (m, 4H), 1.46-1.43 (m, 15H).

Step 2.1-Isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-3-carbonitrile

tert-Butyl3-cyano-1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(70 mg, 0.19 mmol) was dissolved in dichloromethane (3 mL) andtrifluoroacetic acid (0.2 mL) and stirred at ambient temperature for 90minutes. The solvent was concentrated in vacuo and the residue wasco-distilled with toluene followed by ethyl acetate to give 149 mg(100%) of the title compound as a yellow solid: +ESI MS (M+H) 273.5.

Intermediate 14

1-isopropyl-6-methyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one,shown below, was prepared as follows.

Step 1. Benzyl6-bromo-7-hydroxy-1-isopropyl-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl1-isopropyl-1,4-dihydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(4.20 g, 11 mmol), prepared as described in the preparation ofIntermediate 12, was dissolved in 130 mL of tetrahydrofuran. To thereaction was added N-bromosuccinimide (2.49 g, 14 mmol) and 30 mL ofwater. The reaction was stirred for 1 hour at room temperature. Thereaction was partitioned between ethyl acetate and saturated, aqueoussodium chloride. The organic phase was separated then washed anadditional time with saturated aqueous sodium chloride. The organicphase was dried over sodium sulfate and concentrated in vacuo to affordthe title compound as an off-white foam (5.31 g, 100%): +ESI MS (M+H)463.8.

Step 2. Benzyl6-bromo-1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl6-bromo-7-hydroxy-1-isopropyl-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(5.30 g, 11 mmol) was dissolved in 53 mL acetone and then cooled to 0°C. in an ice bath. To the solution was added 83 mL of Jones Reagent(Fillion, E. Tetrahedron Letters 2004, 46, 1091-1094). The ice bath wasremoved and the reaction was allowed to warm to room temperature whereit was stirred for 45 minutes. The reaction was cooled to 0° C. in anice bath and then saturated, aqueous sodium bicarbonate was added untilno further gas evolution was noted. The resulting mixture was filteredthrough a pad of Celite® rinsing with ethyl acetate. The filtrate layerswere separated and the aqueous layer was back extracted with ethylacetate. The organic extracts were combined, washed twice with water,once with saturated, aqueous sodium chloride, dried over sodium sulfateand concentrated in vacuo to afford the title compound (5.27 g, 100%):+ESI MS (M+H) 460.4.

Step 3. Benzyl1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl6-bromo-1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(5.63 g, 12 mmol) was dissolved in 55 mL of acetic acid, to this wasadded zinc powder (2.40 g, 37 mmol). The reaction was stirred for 35minutes at room temperature. The reaction was concentrated in vacuo andthen partitioned between saturated, aqueous sodium bicarbonate and ethylacetate. The phases were separated and the aqueous phase was extractedwith ethyl acetate. The organic extracts were combined, washed withwater, saturated, aqueous sodium chloride, dried over sodium sulfate andconcentrated in vacuo to give an oil. The oil was purified by flashchromatography using 12-100% ethyl acetate in heptane as eluent toafford the title compound as an oil (2.25 g, 48%): +ESI MS(M+H) 382.4;¹H NMR (400 MHz, CDCl₃) δ ppm 7.28-7.40 (m, 6H), 5.32-5.45 (m, 1H), 5.13(s, 2H), 3.41-3.61 (m, 4H), 2.76 (s, 2H), 2.54 (s, 2H), 1.50-1.62 (m,4H), 1.47 (d, J=6.63 Hz, 6H).

Step 4. Benzyl1-isopropyl-6-methyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

Benzyl1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(397 mg, 1.04 mmol) in tetrahydrofuran (8 mL) was cooled to −70° C. Tothis was added lithium bis(trimethylsilyl)amide (1.56 mL, 1.56 mmol) asa 1.0 M solution in tetradhydrofuran over a ten minute period. Theresulting yellow solution was stirred for thirty minutes at −70° C.1,3-Dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidone (1.6 mL) was added tothe reaction, stirring was continued at −70° C. for ten minutes. To thereaction was added iodomethane (746 mg, 5.2 mmol). The reaction wasallowed to warm to room temperature where it was stirred for 18 hours.To the reaction was added saturated, aqueous sodium bicarbonate (2 mL),the mixture was then partitioned between water (20 mL) and ethyl acetate(150 mL). The layers were separated and the aqueous layer was extractedwith ethyl acetate (150 mL). The organic layers were combined, driedover magnesium sulfate, filtered and then concentrated to give a clearoil. The oil was purified by silica gel chromatography using 10-40%ethyl acetate in heptane as eluent to afford the title compound as awhite solid (351 mg, 85%): +ESI MS (M+H) 396.2; ¹H NMR (400 MHz,DMSO-d₆) δ ppm 7.44 (s, 1H), 7.35 (s, 5H), 5.17-5.34 (m, 1H), 5.06 (s,2H), 3.52-3.72 (m, 4H), 2.79 (s, 2H), 2.42-2.48 (m, 1H), 1.38-1.49 (m,4H), 1.35 (t, J=6.74 Hz, 6H), 1.04 (d, J=7.04 Hz, 3H).

Step 5.1-Isopropyl-6-methyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one,shown below, was prepared as follows.

The title compound was prepared from benzyl1-isopropyl-6-methyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylatein an analogous fashion to Intermediate 10, Step 6.

Intermediate 15

1-isopropyl-6,6-dimethyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one,shown below, was prepared as follows.

Step 1. Benzyl 1-isopropyl-6,6-dimethyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate, shown below, was prepared asfollows.

A solution of benzyl1-isopropyl-6-methyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(84 mg, 0.21 mmol), described in the preparation of Intermediate 14, in1 mL tetrahydrofuran was cooled to −70° C. and then treated with lithiumbis(trimethylsilyl)amide (0.318 mL, 0.318 mmol) as a 1.0 M solution intetradhydrofuran over ten minutes. Then1,3-Dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidone (0.2 mL) was added tothe reaction. Stirring continued for ten minutes at −70° C., theniodomethane (152 mg, 1.06 mmol) was added to the reaction. The mixturewas allowed to warm to room temperature where it was held for fourhours. To the reaction was added saturated, aqueous ammonium chloride (1mL), the mixture was then partitioned between water (2 mL) and ethylacetate (10 mL). The layers were separated and the aqueous layer wasextracted with ethyl acetate (5 mL). The organic layers were combined,dried over magnesium sulfate, filtered and then concentrated to give aclear, yellow oil. The oil was purified by silica gel chromatographyusing 10-40% ethyl acetate in heptane as eluent to afford the titlecompound as a clear oil (58 mg, 67%): +ESI MS (M+H) 410.3; ¹H NMR (400MHz, CDCl₃) δ ppm 7.28-7.44 (m, 5H), 7.27 (s, 1H), 5.40 (m, 1H), 5.13(s, 2H), 3.85-4.24 (m, 2H), 2.86-3.11 (m, 2H), 1.58-1.79 (m, 2H), 1.56(s, 2H), 1.46 (d, J=6.64 Hz, 6H), 1.19-1.40 (m, 2H), 1.15 (s, 6H).

Step 2.1-Isopropyl-6,6-dimethyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one,shown below, was prepared as follows.

The title compound was prepared from benzyl1-isopropyl-6,6-dimethyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate in an analogous fashion toIntermediate 10, Step 6.

Intermediate 16

3-bromo-1-tert-butyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one,shown below, was prepared as follows.

Step 1. tert-Butyl1-tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

The hydrochloride salt of Intermediate 10 (1040 mg, 3.492 mmol),di-tert-butyl dicarbonate (800 mg, 3.67 mmol) and triethlyamine (730 mg,7.2 mmol) were combined in dichloromethane (30 mL). The reactionsolution was stirred at ambient temperature for 16 hours. To thereaction was added dichloromethane (20 mL). The reaction solution waswashed with 1N aqueous hydrochloric acid (5 mL), water (5 mL), andsaturated, aqueous sodium chloride (5 mL). The organic phase was driedover magnesium sulfate and concentrated to give tert-butyl1-tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(1262 mg, 100%): −APCI MS (M−H) 360.3; ¹H NMR (400 MHz, CDCl₃) δ ppm7.30 (s, 1H), 3.29-3.56 (m, 4H), 2.77 (s, 2H), 2.56 (s, 2H), 1.67 (s,9H), 1.48-1.56 (m, 4H), 1.46 (s, 9H).

Step 2. tert-Butyl3-bromo-1-tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate,shown below, was prepared as follows.

tert-Butyl1-tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(1090 mg, 3.015 mmol) and sodium acetate (1050 mg, 12.80 mmol) werecombined in ethanol (40 mL) and water (10 mL). To this solution wasadded bromine (1870 mg, 11.7 mmol). The reaction was stirred at roomtemperature for 4 hours. To the reaction was added ethanol (40 mL). Thereaction was stirred for 16 more hours. The reaction solution was pouredin water (20 mL) and extracted twice with ethyl acetate (75 mL each).The combined organic extracts were washed twice with aqueous, saturatedsodium thiosulfate (25 mL each) and saturated, aqueous sodium chloride(25 mL). The organic phase was dried over magnesium sulfate andconcentrated to a final volume of 20 mL to give a precipitate. Themixture was filtered and the solids collected to give the title compoundas a solid (679 mg, 51%): +APCI MS (M+H-Boc) 342.1; ¹H NMR (400 MHz,CDCl₃) δ ppm 3.28-3.60 (m, 4H), 2.66 (s, 2H), 2.56 (s, 2H), 1.65 (s,9H), 1.48-1.55 (m, 4H), 1.46 (s, 9H).

Step 3.3-Bromo-1-tert-butyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one,shown below, was prepared as follows.

tert-Butyl3-bromo-1-tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-carboxylate(670 mg, 1.52 mmol) and 4 M hydrogen chloride in dioxane (8 mL) werecombined and stirred for 2.5 hours. To the reaction was added diethylether (20 mL). A precipitate formed that was filtered and the solidscollected to give the title compound (573 mg, 97%): +APCI MS (M+H)342.1; ¹H NMR (400 MHz, CD₃OD) δ ppm 3.24 (t, J=5.96 Hz, 4H), 2.80 (s,2H), 2.74 (s, 2H), 1.71-1.92 (m, 4H), 1.65 (s, 9H).

Intermediate 17

1′-Isopropyl-1′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-one,shown below, was prepared as follows.

Step 1. 1-(4-Hydroxy-1-(4-methoxybenzyl)-1H-pyrazol-3-yl)ethanone, shownbelow, was prepared as follows:

To a stirred solution of (4-methoxybenzyl)hydrazine hydrochloride (13.5g, 71.5 mmol) in water (400 mL) was added a solution of pyruvaldehyde(5.2 g, 71.5 mmol) in water (200 mL) over a 10-min period. After anadditional 50 min, the reaction mixture was extracted withdichloromethane (3×300 mL), the combined extracts dried over sodiumsulfate, concentrated in vacuo and the residue was used in the nexttransformation without further purification.

A stirred solution of the product from the above reaction (12.3 g, 59.8mmol) and glyoxal (43 g, 299 mmol) in methanol (34 mL)/water (300 mL)was heated at 100° C. for 5 h. The reaction mixture was cooled, dilutedwith ethyl acetate, the organic phase dried over sodium sulfate andconcentrated in vacuo. Purification of the residue was performed on anCombiflash® unit (300 g column, gradient 10-35% ethyl acetate:heptanes)afforded the title compound (6.04 g). ¹H NMR (400 MHz, CDCl₃) δ ppm 8.05(s, 1H) 7.18 (m, J=8.79 Hz, 2H), 6.93 (s, 1H), 6.87 (m, J=8.60 Hz, 2H),5.15 (s, 2H), 3.79 (s, 3H), 2.55 (s, 3H); m/z (M+1)=247.0.

Step 2.1-Benzyl-2′-(4-methoxybenzyl)-2′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-one,shown below, was prepared as follows.

Suspended 1-(4-hydroxy-1-(4-methoxybenzyl)-1H-pyrazol-3-yl)ethanone (350mg, 1.42 mmol) in 10 mL methanol and added N-benzyl-4-piperidone (0.25mL, 1.42 mmol) and pyrrolidine (0.036 mL, 0.3 eq). The mixture was thenheated at reflux for 18 h. The reaction was then cooled to roomtemperature and methanol was removed under reduced pressure. Theresultant orange oil was partitioned between 50 mL ethyl acetate and 50mL water. The aqueous phase was extracted with an additional 50 mL ethylacetate. The organic layers were combined and dried over sodium sulfate,filtered and concentrated. The resultant oil was flash chromatographed(50-100% ethyl acetate/heptane gradient, 25 g silica) to yield 428 mg(72%) of1-benzyl-2′-(4-methoxybenzyl)-2′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-oneas a pale yellow solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.25-7.37 (m, 6H),6.95 (s, 1H), 6.81-6.92 (m, 2H), 5.20 (s, 2H), 3.79 (s, 3H), 3.49 (s,2H), 2.63 (s, 2H), 2.57 (d, J=11.3 Hz, 2H), 2.25-2.44 (m, 2H), 2.02 (d,J=12.5 Hz, 2H), 1.62-1.77 (m, 2H); m/z (M+1)=418.5.

Step 3.1-Benzyl-1′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-one,shown below, was prepared as follows.

Dissolved1-benzyl-2′-(4-methoxybenzyl)-2′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-one(428 mg, 1.02 mmol) in 20 mL 1,2-dichloroethane and treated with 10 mLtrifluoroacetic acid. The resultant mixture was heated for 18 h at 90°C. The reaction was cooled to ambient temperature and concentrated todryness under reduced pressure. The resultant residue was taken up in 50mL saturated aqueous sodium bicarbonate and extracted with 2×50 mL ethylacetate. The organic extracts were combined and dried over sodiumsulfate, filtered and concentrated under reduced pressure. The resultantoil was flash chromatographed (25 g silica, stepgradient 5 columnvolumes 50% ethyl acetate/heptane, 10 CV 100% ethyl acetate) to yield278 mg (91%) of1-benzyl-1′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-one asa pale yellow solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 7.27 (m, 6H), 3.55(s, 2H), 2.64 (m, 4H), 2.45 (td, J=11.7, 2.5 Hz, 2H), 2.06 (d, J=12.1Hz, 2H), 1.74 (m, 2H); m/z (M+1)=298.5.

Step 4.1-Benzyl-t-isopropyl-1′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-one,shown below, was prepared as follows.

1-Benzyl-1′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-one(204 mg, 0.67 mmol) was dissolved in 10 mL tetrahydrofuran. 2-propanol(0.11 mL, 1.37 mmol) and polymer supported triphenylphosphine (0.5 g, 3mmol/g loading) were added followed by addition of DBAD (322 mg, 1.37mmol) and stirred at ambient temperature for 5 days. Filtered off poymersupported triphenyl phosphine and washed filtercake with 100 mL ethylacetate. The filtrate was concentrated and the resultant yellow oil wastreated with 10 mL 4N HCl/dioxane. The mixture was stirred 1 h atambient temperature. The volatiles were removed under reduced pressure.The resultant sludge was partitioned between 50 mL sat aq. sodiumbicarbonate and 50 mL ethyl acetate. The organic phase was dried oversodium sulfate, filtered and concentrated under reduced pressure. Theresultant oil was flash chromatographed (30-100% ethyl acetate/heptanegradient, 10 g silica) to yield 90 mg (35%) of1-benzyl-1′-isopropyl-1′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-oneas a pale yellow solid. ¹H NMR (400 MHz, CDCl₃) δ ppm 7.30 (m, 3H), 7.24(m, 2H), 7.18 (s, 1H), 5.14 (spt, 1H), 3.52 (s, 2H), 2.60 (m, 4H), 2.41(td, J=11.6, 2.5 Hz, 2H), 2.06 (d, J=12.5 Hz, 2H), 1.71 (m, 2H), 1.44(m, 6H): m/z (M+1)=340.5.

Step 5.1′-Isopropyl-1′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-one,shown below, was prepared as follows.

1-Benzyl-1′-isopropyl-1′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-one(81 mg, 0.22 mmol) was dissolved in 10 mL 1,2-dichloroethane. Added1-chloroethyl chloroformate (60 mL, 0.54 mmol) was added and the mixturewas stirred 1 h at reflux then cooled to room temperature. The volatileswere removed under reduced pressure and the residue taken up in 10 mLmethanol and heated at reflux for 1 h. The mixture was cooled to ambienttemperature and the volatiles were removed under reduced pressure. Theresidue was taken up in 50 mL saturated aqueous sodium bicarbonate andextracted 2×30 mL ethyl acetate. The combined organic extracts weredried over sodium sulfate, filtered and concentrated to yield 46 mg(86%) of1′-isopropyl-1′H-spiro[piperidine-4,5′-pyrano[3,2-c]pyrazol]-7′(6′H)-oneas a white solid. ¹H NMR (400 MHz, CD₃OD) δ ppm 7.20 (s, 1H), 5.13 (spt,1H), 2.93 (m, 2H), 2.80 (dt, J=12.8, 4.0 Hz, 2H), 2.64 (s, 2H), 2.02 (m,2H), 1.63 (m, 2H), 1.40 (m, 6H); m/z (M+1)=250.2

Example 1

4-((4-(1-Tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)phenoxy)methyl)benzoicacid, shown below, was prepared as follows.

Step 1. Methyl4-((4-(1-tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)phenoxy)methyl)benzoate,shown below was prepared as follows.

A solution of 4-(4-(methoxycarbonyl)benzyloxy)benzoic acid (25 mg, 0.087mmol), 1-tert-butyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one(26 mg, 0.087 mmol), diisopropylethylamine (53 μL, 0.30 mmol),1,2,3-benzotriazole-1-ol, monohydrate (14 mg, 0.087 mmol),4-dimethylaminopyridine (1.1 mg, 0.01 mmol) and1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (19 mg, 0.96 mmol) indichloromethane (0.2 mL) was heated at 30° C. for 18 hr. The reactionmixture was partitioned between ethyl acetate/aqueous 0.3N hydrochloricacid, the organic phase was dried over sodium sulfate, concentrated invacuo and purified by preparative HPLC to afford methyl4-((4-(1-tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)phenoxy)methyl)benzoate. HPLC column: Waters Atlantis C18 4.6×50 mm, 5 um, solvent:acetonitrile:water (0.05% TFA); flow rate 2 mL/min; gradient (% organic)start=5%, end=95%, gradient time=4 min, retention time=3.68 min; m/z=530(M+1).

Step 2. To a stirred solution of methyl4-((4-(1-tert-butyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)phenoxy)methyl)benzoate(39 mg, 0.07 mmol) in tetrahydrofuran (0.75 mL) was added aqueouslithium hydroxide (0.22 mL, 0.22 mmol). After 18 hr, 0.3 mL of 1Naqueous hydrochloric acid, and 0.2 mL of saturated aqueous sodiumchloride were added. The resulting mixture was extracted with2-methyltetrahydrofuran (3×4 mL), the combined organic layers dried oversodium sulfate and concentrated in vacuo to afford a gum (46 mg).Purification by preparative HPLC afforded the title compound (26 mg).HPLC column: Waters Atlantis C18 4.6×50 mm, 5 um, solvent:acetonitrile:water (0.05% TFA); flow rate 2 mL/min; gradient (% organic)start=5%, end=95%, gradient time=4 min, retention time=3.19 min; m/z=516(M+1).

Example 2

3-(4-(1-Isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)-6-methoxypyridin-2-yl)benzoicacid, shown below, was prepared as follows.

Step 1. Tert-butyl3-(4-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)-6-methoxypyridin-2-yl)benzoate,shown below, was prepared as follows.

A solution of 2-(4-(tert-butoxycarbonyl)phenyl)-6-methoxyisonicotinicacid (500 mg, 1.52 mmol) and 1,1-carbonyldiimidazole (271 mg, 1.67 mmol)in tetrahydrofuran (30 mL) was stirred at reflux for 1 hr. After coolingto room temperature1-isopropyl-4,6-dihydrospiro[indazole-5,4′-piperidin]-7(1H)-one (539 mg,1.68 mmol) and triethylamine (0.32 mL, 2.28 mmol) were addedsequentially and the resulting slurry was heated at reflux temperaturefor 1 hr. After cooling, the reaction mixture was diluted into ethylacetate, washed with 1N aqueous hydrochloric acid, saturated aqueoussodium chloride, dried over sodium sulfate to afford tert-butyl3-(4-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)-6-methoxypyridin-2-yl)benzoate(850 mg) as a white foam. This material was taken onto the next stepwithout further purification. ¹H NMR (400 MHz, CDCl₃) δ ppm 8.60 (t,J=1.66 Hz, 1H), 8.20 (dt, J=8.05, 1.44 Hz, 1H), 8.01 (dt, J=7.80, 1.37Hz, 1H), 7.50 (t, J=7.80 Hz, 1H), 7.36 (s, 1H), 7.33 (d, J=0.98 Hz, 1H),6.64 (d, 1H), 5.35 (spt, J=6.44 Hz, 1H), 4.04 (s, 3H), 3.71-3.84 (m,2H), 3.36-3.42 (m, 2H), 2.79 (d, J=2.34 Hz, 2H), 2.58 (s, 2H), 1.66-1.72(m, 2H), 1.61 (s, 9H), 1.56 (s, 2H), 1.40-1.48 (m, 6H); m/z (M+1)=559.2.

Step 2. A solution of tert-butyl3-(4-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]1′-ylcarbonyl)-6-methoxypyridin-2-yl)benzoate(850 mg, 1.52 mmol) in dichloromethane (40 mL) and trifluoroacetic acid(13 mL) was stirred for 18 hr. The solvents were removed in vacuo andthe residue was purified by flash chromatography (40 g silica gelcolumn, eluting with a gradient of 20-100% ethyl acetate:heptanes (0.5%acetic acid) to afford the title compound as a white solid (568 mg). ¹HNMR (400 MHz, CD₃OD) δ ppm 8.72 (t, J=1.66 Hz, 1H), 8.31 (dt, J=8.15,1.39 Hz, 1H), 8.06 (dt, J=7.80, 1.27 Hz, 1H), 7.57 (t, J=7.80 Hz, 1H),7.48 (d, J=0.98 Hz, 1H), 7.40 (s, 1H), 6.74 (d, 1H), 5.36 (spt, J=6.76Hz, 1H), 4.05 (s, 3H), 3.82-3.91 (m, 1H), 3.65-3.74 (m, 1H), 3.44 (t,J=5.76 Hz, 2H), 3.24 (s, 0H), 2.87 (d, J=1.37 Hz, 2H), 2.63 (d, J=2.93Hz, 2H), 1.63-1.71 (m, 2H), 1.52-1.60 (m, 2H), 1.41 (d, J=6.24 Hz, 3H),1.38 (d, J=6.24 Hz, 3H); m/z (M+1)=503.2

Example 3

3-(4-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)-6-oxo-1,6-dihydropyridin-2-yl)benzoicacid, shown below, was prepared as follows.

A mixture of3-(4-(1-isopropyl-7-oxo-1,4,6,7-tetrahydrospiro[indazole-5,4′-piperidine]-1′-ylcarbonyl)-6-methoxypyridin-2-yl)benzoicacid (30 mg, 0.06 mmol), potassium iodide (30 mg, 0.18 mmol) in aceticacid (1 mL) was stirred at 120° C. for 7 hr. The reaction mixture wasfiltered, concentrated in vacuo and purified by preparative HPLC toafford the title compound (12 mg). HPLC column: Waters Atlantis C184.6×50 mm, 5 um, solvent: acetonitrile:water (0.05% TFA); flow rate 2mL/min; gradient (% organic) start=5%, end=95%, gradient time=4 min,retention time=2.32 min; m/z=489.1451 (M+1).

The compounds listed in Table 1 below were prepared using proceduresanalogous to those described above for the synthesis of the compounds ofExamples 1-3 using the appropriate starting materials which areavailable commercially, prepared using preparations well-known to thoseskilled in the art, or prepared in a manner analogous to routesdescribed above for other intermediates. The compounds listed below wereisolated initially as the free base and may be converted to apharmaceutically acceptable salt for testing.

TABLE 1

Ex. R¹ —C(O)—A¹—L—A² Analytical Data  4 t-butyl

MS (MH + 1) 516.3, ¹H NMR (CD₃OD, 400 MHz): 8.70 (s, 1H), 8.27 (d, 1H),8.03 (d, 1H), 7.46 (t, 1H), 7.45 (d, 1H), 7.31 (s, 1H), 7.16 (d, 1H),3.40-3.75 (br.s, 4H), 3.05 (s, 3H), 2.86 (s, 2H), 2.60 (s, 2H), 1.61 (s,13H).  5 isopropyl

MS (MH + 1) 558.3; ¹H NMR (CDCl₃, 400 MHz): 8.36 (s, 1H), 8.27 (d, 1H),7.49 (t, 1H), 7.37 (s, 2H), 7.05 (d, 1H), 3.56 (h, 1H), 3.61 (m, 4H),3.07 (s, 3H), 2.58 (s, 2H), 1.48 (m, 4H), 1.44 (d, 6H).  6 t-butyl

MS (MH + 1) 486.24; ¹H NMR (400 MHz, CDCl₃) 8.30 (s, 1H), 8.21 (d, 1H),7.79 (d, 1H), 7.65 (d, 2H), 7.55 (t, 1H), 7.50 (d, 2H), 7.31 (s, 1H),3.62 (br, 1H), 3.48 (br, 2H), 2.82 (s, 2H), 2.62 (s, 2H), 1.80-1.48 (m,13H), 0.86 (br, 1H).  7 isopropyl

MS (MH + 1) 472.26; ¹H NMR (400 MHz, CDCl₃) 8.31 (s, 1H), 8.09 (d, 1H),7.79 (d, 1H), 7.65 (d, 2H), 7.53 (t, 1H), 7.50 (d, 2H), 7.38 (s, 1H),5.37-5.35 (m, 1H), 3.82 (br, 1H), 3.47 (br, 2H), 2.81 (s, 2H), 2.60 (s,2H), 1.70-1.55 (m, 5H), 1.45 (d, 6H).  8 isopropyl

MS (MH + 1) 502.27; ¹H NMR (CD₃OD, 400 MHz) δ ppm 8.15 (d, 2H), 8.07 (d,2H), 7.47 (d, 1H), 7.40 (s, 1H), 7.19 (d, 1H), 5.35 (h, 1H), 3.50 (br.s, 4H), 3.05 (s, 3H), 2.86 (s, 2H). 2.62 (s, 2H), 1.60 (br. s, 4H), 1.40(d, 6H).  9 isopropyl

MS (MH + 1) 503.5; ¹H NMR (400 MHz, CDCl₃) δ ppm 1.39- 1.48 (m, 6 H),1.53 (bs, 2H), 1.70 (br. s., 2H), 2.59 (s, 2 H), 2.80 (s, 2H), 3.40 (br.s., 2 H), 3.71-3.88 (m, 2H), 4.06 (s, 3 H) 5.31-5.42 (m, 1 H) 6.68 (s, 1H), 7.38 (s, 2 H), 8.15 (m, 4 H). 10 t-butyl

MS (MH + 1) 517.6; ¹H NMR (400 MHz, CDCl₃) δ ppm 1.50- 1.57 (m, 2H),1.66 (s, 9 H), 1.67-1.74 (m, 2H), 2.64 (s, 2 H) 2.83 (s, 2 H), 3.39-3.38(m, 2H), 3.72-3.91 (m, 4H), 4.07 (s, 3 H) 6.71 (d, J = 0.98 Hz, 1 H)7.32 (s, 1 H) 7.39 (d, J = 0.98 Hz, 1 H) 8.05-8.26 (m, 4 H) 11 t-butyl

MS (M + H) 517.6; 1H NMR (400 MHz, CDCl₃) δ ppm 1.49- 1.57 (m, 2H), 1.66(s, 9 H), 1.68-1.75 (m, 2H), 2.64 (s, 2 H), 2.84 (s, 2 H), 3.43 (bs,2H), 3.72-3.92 (m, 2H), 4.08 (s, 3 H), 6.68 (d, J = 0.98 Hz, 1 H), 7.32(s, 1 H), 7.45 (d, J = 0.98 Hz, 1 H), 7.59 (t, 1 H), 8.16 (d, J = 7.82Hz, 1 H), 8.34 (d, J = 7.82 Hz, 1 H), 8.76 (s, 1 H). 12 t-butyl

MS (MH + 1) 406.5; ¹H NMR (400 MHz, CDCl₃) δ ppm 8.00 (s, 1 H) 7.79 (s,1 H) 7.32- 7.47 (m, 3 H) 5.30-5.41 (m, 1 H) 4.07 (s, 3 H) 3.61 (br. s.,4 H) 2.79 (s, 2 H) 2.58 (s, 2 H) 1.61 (br. s., 2 H) 1.53 (br. s., 2 H)1.43 (d, J = 6.84 Hz, 6 H) 13 t-butyl

MS (MH + 1) 516.28; ¹H NMR (CDCl₃, 400 MHz): 8.16 (m, 4H), 7.38 (br.s,2H), 7.05 (d, 1H), 5.37 (h, 1H), 3.48 (m 6H), 2.60 (s, 2H), 2.16 (br.s,4H), 1.45 (d, 6H), 1.23 (app. br.s, 3H). 14 isopropyl

MS (MH + 1) 511, ¹H NMR (CDCl₃, 400 MHz) δ ppm 8.16 (m, 4H), 7.38 (br.s,2H), 7.05 (d, 1H), 5.37 (h, 1H), 3.48 (m 6H), 2.60 (s, 2H), 2.16 (br.s,4H), 1.45 (d, 5H), 1.23 (app. br.s, 3H). 15 isopropyl

MS (MH + 1) 511; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 13.04 (br. s., 1 H),11.77 (s, 1 H), 8.16 (s, 1 H), 7.93 (d, J = 7.8 Hz, 1 H), 7.89 (d, J =7.8 Hz, 1 H), 7.60 (t, J = 7.7 Hz, 1 H), 7.40-7.48 (m, 2 H), 7.27 (t, J= 7.7 Hz, 1 H), 7.14 (d, J = 7.2 Hz, 1 H), 6.74 (d, J = 1.2 Hz, 1 H),5.24 (spt, J = 6.6 Hz, 1 H), 3.75 (br. s., 2 H), 3.65 (br. s., 2 H),2.79 (s, 2 H), 2.58 (s. 2 H), 1.50 (br. s., 4H), 1.33 (d, J = 6.6 Hz, 6H). 16 isopropyl

MS (MH + 1) 511; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.75-13.11 (m, 1 H),11.78 (s, 1 H), 8.03 (d, J = 8.4 Hz, 2 H), 7.76 (d, J = 8.2 Hz, 2 H),7.40- 7.50 (m, 2 H), 7.27 (t, J = 7.7 Hz, 1 H), 7.17 (d, J = 7.0 Hz, 1H), 6.79 (d, J = 1.2 Hz, 1 H), 5.25 (spt, J = 6.5 Hz, 1 H), 3.76 (br.s., 2H), 3.66 (br. s., 2 H), 2.79 (s, 2H), 2.59 (s, 2 H), 1.44- 1.58 (m,4 H), 1.33 (d, J = 6.6 Hz, 6 H). 17 t-butyl

MS (MH + 1) 525; ¹H NMR (400 MHz, DMSO-d₆) ^(d) ppm 11.76 (s, 1 H), 8.16(s, 1 H), 7.93 (d, J = 7.8 Hz, 1 H), 7.88 (d, J = 7.4 Hz, 1 H), 7.59 (t,J = 7.7 Hz, 1 H), 7.43 (d, J = 8.2 Hz, 1 H), 7.36 (s, 1 H), 7.27 (t, J =7.7 Hz, 1 H), 7.14 (d, J = 7.2 Hz, 1 H), 6.74 (s, 1 H), 3.78 (d, J =12.9 Hz, 2H), 3.63 (br. s., 2 H), 2.81 (s, 2 H), 2.60 (s, 2 H), 1.55 (s.9 H), 1.49 (br. s., 4 H) 18 t-butyl

MS (MH + 1); ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.77 (s, 1 H), 8.03 (d, J= 8.2 Hz, 2 H), 7.76 (d, J = 8.2 Hz, 2 H), 7.45 (d, J = 8.2 Hz, 1 H),7.37 (s, 1 H), 7.27 (t, J = 7.7 Hz, 1 H), 7.16 (d, J = 7.2 Hz, 1 H),6.79 (d, J = 1.0 Hz, 1 H), 3.78 (br. s., 2 H), 3.64 (br. s., 2 H), 2.82(s, 2 H), 2.61 (s, 2 H), 1.55 (s, 9H), 1.49 (br. s., 4 H). 19 t-butyl

MS (MH + 1) 530.31; ¹H NMR (CD₃OD, 400 MHz) δ ppm 8.64 (s, 1H), 8.25 (d,1H), 8.02 (d, 1H), 7.80 (br. s, NH), 7.50 (m, 2H), 7.30 (s, 1H), 7.10(d, 1H). 20 isopropyl

MS (MH + 1) 516.28; ¹H NMR (DMSO-d₆, 400 MHz) δ ppm 8.15 (d, 2H), 7.97(d, 2H), 7.41 (d, 1H), 7.36 (s, 1H), 7.20 (d, 1H), 6.23 (br.s, NH), 3.87(br.s, 4H), 2.92 (s, 3H), 2.78 (s, 2H), 2.58 (s, 2H), 1.54 (s, 9H), 1.46(br.s, 4H). 21 t-butyl

MS (MH + 1) 477; LC/MS retention time 2.83 minutes on a Waters AtlantisdC18 4.6 × 50 mm, 5 μm gradient elution (5% to 95%) with water:acetonitrile (0.05% TFA), 4 minute gradient and 5 minute hold time 22isopropyl

MS (MH + 1) 463; LC/MS retention time 2.57 minutes on a Waters AtlantisdC18 4.6 × 50 mm, 5 μm gradient elution (5% to 95%) with water:acetonitrile (0.05% TFA), 4 minute gradient and 5 minute hold time 23isopropyl

MS (MH + 1) 530.29; ¹H NMR (CDCl₃, 400 MHz) δ ppm 8.72 (s, 1H), 8.33 (d,1H), 8.12 (d, 1H), 7.52 (t, 1H), 7.36 (d, 2H), 7.05 (d, 1H), 5.67 (br.s, 1H), 5.36 (h, 1H), 4.43 (m, 1H), 3.62 (m, 4H), 3.09 (s, 1H), 2.81 (s,2H), 2.59 (s, 2H), 1.61 (m, 4H), 1.45 (d, 6H), 1.30 (d, 6H) 24 t-butyl

MS (MH + 1) 544.3; ¹H NMR (CDCl₃, 400 MHz) δ ppm 8.57 (s, 1H), 8.36 (d,1H), 7.36-7.54 (m, 2H), 7.31 (s, 1H), 7.04 (d, 1H), 4.22 (br. s, 1H),3.65 (br.s, 4H), 2.83 (s, 2H), 2.62 (s, 2H), 1.65 (s, 13 H), 1.32 (d,6H) 25 isopropyl

MS (MH + 1) 530.29; ¹H NMR (CDCl₃, 400 MHz) δ ppm 8.10- 8.18 (dd, 4H),7.38 (m, 2H), 7.04 (d, 1H), 5.39 (h, 1H), 4.36 (br. s, 1H), 3.51 (br. s,4H), 2.81 (s, 2H), 2.59 (s, 2H), 1.70 (m, 4H), 1.45 (d, 6H), 1.29 (d,6H) 26 isopropyl

MS (MH + 1) 512.3; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.24 (s, 1 H), 8.07(m, 2 H), 7.86 (m, 2 H), 7.57 (s, 1 H), 7.43 (s, 1 H), 7.25 (d, J = 0.98Hz, 1 H), 5.24 (t, J = 6.63 Hz, 1 H), 3.71 (br. s., 2 H), 3.59 (br. s.,2H), 2.78 (br. s., 2 H), 2.60 (s, 2 H), 1.46 (br. s., 4 H), 1.32 (d). 27isopropyl

MS (MH + 1) 489.15; LC/MS retention time 2.29 minutes on a WatersAtlantis dC18 4.6 × 50 mm, 5 

 m gradient elution (5% to 95%) with water: acetonitrile (0.05% TFA), 4minute gradient and 5 minute hold time 28 isopropyl

MS (MH + 1) 512.2; ¹H NMR (400 MHz, CD₃OD) δ ppm 8.54 (d, J = 6.06 Hz, 1H), 8.48 (t, J = 1.66 Hz, 1 H), 8.25 (dt, J = 7.77, 1.39 Hz, 1 H), 8.10(dt, J = 8.11, 1.42 Hz, 1 H), 7.70- 7.80 (m, 2 H), 7.41 (s, 1 H), 7.17(s, 1 H), 5.32-5.42 (m, 1 H), 3.85 (br. s., 4 H), 2.89 (s, 2 H), 2.64(s, 2 H), 1.66 (br. s., 4 H) 1.41(d). 29 isopropyl

MS (MH + 1) 512.3; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 8.37 (d, J = 4.88 Hz,1 H) 8.07 (m, 2 H) 7.87 (m, 2 H) 7.43 (s, 1 H) 7.26 (d, J = 4.88 Hz, 1H) 6.79 (d, J = 2.15 Hz, 1 H) 5.23 (d, J = 6.44 Hz, 1 H) 3.69 (s, 4 H)2.78 (s, 2 H) 2.59 (s, 2 H) 1.50 (br. s., 4 H) 1.32 (d). 30 isopropyl

MS (MH + 1) 512; ¹H NMR (400 MHz, CD₃OD) δ ppm 8.46 (d, J = 6.8 Hz, 1H), 8.33-8.37 (m, 2 H), 8.05-8.10 (m, 2 H), 7.99 (dd, J = 6.7, 0.7 Hz, 1H), 7.43 (s, 1 H), 7.34 (d, J = 0.8 Hz, 1 H), 5.38 (spt, J = 6.5 Hz, 1H), 3.75-3.94 (m, 4 H), 2.91 (s, 2 H), 2.66 (s, 2 H), 1.68 (br. s., 4H), 1.43 (s, 6 H). 31 isopropyl

MS (MH + 1) 555.3; LC/MS retention time 3.09 minutes. Column: WatersAtlantis dC18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/ 95% MeCN to5.0 min. Flow: 2.0 mL/min. 32 isopropyl

MS (MH + 1) 516.3; LC/MS retention time 2.77 minutes. Column: WatersAtlantis dC18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/95% MeCN to5.0 min. Flow: 2.0 mL/min. 33 isopropyl

MS (MH + 1) 516; ¹H NMR (400 MHz, CD₃OD) δ ppm 8.18 (d, J = 8.4 Hz, 2H), 7.97 (d, J = 8.4 Hz, 2H), 7.42 (s, 1 H), 7.15 (d, J = 0.8 Hz, 1 H),7.07 (s, 1 H), 5.38 (spt, J = 6.7 Hz, 1 H), 3.83- 3.91 (m, 1 H),3.69-3.76 (m, 1 H), 3.45-3.52 (m, 2 H), 3.33- 3.36 (m, 6 H), 2.90 (s, 2H), 2.66 (d, J = 2.7 Hz, 2 H), 1.68- 1.74 (m, 2 H), 1.58-1.64 (m, 2 H),1.42 (t, J = 6.3 Hz, 6 H). 34 isopropyl

MS (MH + 1) 513.3; LC/MS retention time 2.4 minutes. Column: WatersAtlantis dC18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/95% MeCN to5.0 min. Flow: 2.0 mL/min. 35 isopropyl

MS (MH + 1) 512.2; LC/MS retention time 2.2 minutes. Column: WatersAtlantis dC18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/95% MeCN to5.0 min. Flow: 2.0 mL/min. 36 isopropyl

MS (MH + 1) 513.3; LC/MS retention time 2.41 minutes. Column: WatersAtlantis dC18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/ 95% MeCN to5.0 min. Flow: 2.0 mL/min. 37 isopropyl

MS (MH + 1) 511.2; LC/MS retention time 2.82 minutes. Column: WatersAtlantis dC18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/ 95% MeCN to5.0 min. Flow: 2.0 mL/min. 38 isopropyl

MS (MH + 1) 512.24; LC/MS retention time 2.21 minutes. Column: WatersAtlantis dC18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/ 95% MeCN to5.0 min. Flow: 2.0 mL/min. 39 isopropyl

MS (MH + 1) 511.3; LC/MS retention time 2.87 minutes. Column: WatersAtlantis dC18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/ 95% MeCN to5.0 min. Flow: 2.0 mL/min. 40 isopropyl

MS (MH + 1) 512.28; LC/MS retention time 2.27 minutes. Column: WatersAtlantis dC18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/ 95% MeCN to5.0 min. Flow: 2.0 mL/min. 41 isopropyl

MS (MH + 1) 511.2; LC/MS retention time 3.19 minutes. Column: WatersAtlantis dC18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/ 95% MeCN to5.0 min. Flow: 2.0 mL/min. 42 isopropyl

MS (MH + 1) 570.27; LC/MS retention time 3.11; minutes. Column: WatersAtlantis dC18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/ 95% MeCN to5.0 min. Flow: 2.0 mL/min. 43 isopropyl

MS (MH + 1) 502; LC/MS retention time 2.06 minutes. Column: WatersSunfire C18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/ 95% MeCN to5.0 min. Flow: 2.0 mL/min. 44 isopropyl

MS (MH + 1) 502; LC/MS retention time 2.09 minutes. Column: WatersSunfire C18 4.6 × 50 mm, 5 um Modifier: TFA 0.05% Gradient: 95% H20/5%MeCN linear to 5% H20/95% MeCN over 4.0 min, HOLD at 5% H20/ 95% MeCN to5.0 min. Flow: 2.0 mL/min. 45 isopropyl

MS (MH + 1) 516; ¹H NMR (CD₃OD + 1 drop DCM, 400 MHz): 8.70 (s, 1H),8.14 (d, 1H), 8.03 (d, 1H), 7.59 (m, 2H), 7.50 (m, 1H), 7.42 (d, 1H),7.41 (s, 1H), 5.41 (m, 1h), 4.45 (m, 1H), 4.28 (m, 1H), 3.89 (m, 1H),3.73 (m, 1h), 2.89 (dd, 2H), 2.63 (s, 2H), 1.79 to 1.62 (m,4H), 1.42 (d,6H). 46 isopropyl

MS (M + H) 512; ¹H NMR (400 MHz, CD₃OD) δ ppm 8.09 (d, J = 8.2 Hz, 2 H),7.77 (d, J = 8.6 Hz, 2 H), 7.66 (m, 1 H), 7.43 (s, 1 H), 5.39 (spt, J =6.6 Hz, 1 H), 4.30-4.39 (m, 1 H), 4.14- 4.22 (m, 1 H), 3.86-3.95 (m, 1H), 3.70-3.79 (m, 1 H), 2.91 (s, 2H), 2.66 (s, 2 H), 1.66- 1.74 (m, 4H), 1.42 (d, J = 6.8 Hz, 6 H) 47 isopropyl

MS (M + H) 512; ¹H NMR (400 MHz, CD₃OD) δ ppm 8.30 (t, J = 1.7 Hz, 1 H),7.99 (d, J = 7.8 Hz, 1 H), 7.87 (d, J = 7.8 Hz, 1 H), 7.59-7.66 (m, 1H), 7.54 (t, J = 7.8 Hz, 1 H), 7.41 (s, 1 H), 5.38 (spt, J = 6.6 Hz, 1H), 4.30-4.40 (m, 1 H), 4.13- 4.23 (m, 1 H), 3.85-3.95 (m, 1 H),3.69-3.79 (m, 1 H), 2.90 (s, 2H), 2.65 (s, 2 H), 1.63- 1.77 (m, 4 H),1.42 (dd, J = 6.6, 1.8 Hz, 6 H) 48 isopropyl

MS (MH + 1) 496; ¹H NMR (CDCl₃, 400 MHz): 8.31 (s, 1H), 8.02 (d, 1H),7.78 (m, 3H), 7.65 (m, 1H), 7.50 (m, 2H), 7.41 (s, 1H), 5.37 (m, 1H),3.86 (br.s, 1H), 3.72 (br.s, 1H), 2.88 (s, 2H), 2.50 (s, 2H), 1.60 (m,4H), 1.40 (d, 6H) 49 t-butyl

MS (MH + 1) 510.25; ¹H NMR (CDCl₃,400 MHz): 8.32 (s, 1H), 8.00 (s, 1H),7.80 (m, 3H), 7.60 (t, 1H), 7.54 (d, 2H), 7.32 (s, 1H), 3.90 (br.s, 1H),3.70 (br.s, 1H), 3.50 (br.s, 2H), 2.85 (s, 2H), 2.65 (s, 2H), 1.56 (m,13 H).

Pharmacological Data Biological Protocols

The utility of the compounds of present invention, in the treatment ofdiseases (such as are detailed herein) in animals, particularly mammals(e.g., humans) may be demonstrated by the activity thereof inconventional assays known to one of ordinary skill in the art, includingthe in vitro and in vivo assays described below. Such assays alsoprovide a means whereby the activities of the compound of the presentinvention can be compared with the activities of other known compounds.

Direct Inhibition of the Activities of ACC1 and ACC2

The ACC inhibitory activity of the compound of the present invention wasdemonstrated by methods based on standard procedures. For example directinhibition of ACC activity, for the compound of Formula (1) wasdetermined using preparations of recombinant human ACC1 (rhACC1) andrecombinant human ACC2 (rhACC2). Representative sequences of therecombinant human ACC1 and ACC2 that can be used in the assay areprovided in FIG. 1 (SEQ ID NO. 1) and FIG. 2 (SEQ. ID NO. 2),respectively.

[1] Preparation of rhACC1. Two liters of SF9 cells, infected withrecombinant baculovirus containing full length human ACC1 cDNA, weresuspended in ice-cold lysis buffer (25 mM Tris, pH 7.5; 150 mM NaCl; 10%glycerol; 5 mM imidazole (EMD Bioscience; Gibbstown, N.J.); 2 mM TCEP(BioVectra; Charlottetown, Canada); Benzonase nuclease (10000 U/100 gcell paste; Novagen; Madison, Wis.); EDTA-free protease inhibitorcocktail (1 tab/50 mL; Roche Diagnostics; Mannheim, Germany). Cells werelysed by 3 cycles of freeze-thaw and centrifuged at 40,000×g for 40minutes (4° C.). Supernatant was directly loaded onto a HisTrap FF crudecolumn (GE Healthcare; Piscataway, N.J.) and eluted with an imidazolegradient up to 0.5 M over 20 column volumes (CV). ACC1-containingfractions were pooled and diluted 1:5 with 25 mM Tris, pH 7.5, 2 mMTCEP, 10% glycerol and direct loaded onto a CaptoQ (GE Healthcare)column and eluted with an NaCl gradient up to 1 M over 20 CV's.Phosphate groups were removed from purified ACC1 by incubation withlambda phosphatase (100 U/10 μM target protein; New England Biolabs;Beverly, Mass.) for 14 hours at 4° C.; okadaic acid was added (1 μMfinal concentration; Roche Diagnostics) to inhibit the phosphatase.Purified ACC1 was exchanged into 25 mM Tris, pH 7.5, 2 mM TCEP, 10%glycerol, 0.5 M NaCl by 6 hour dialysis at 4° C. Aliquots were preparedand frozen at −80° C.

[2] Measurement of rhACC1 inhibition. hACC1 was assayed in a Costar#3676 (Costar, Cambridge, Mass.) 384-well plate using the TranscreenerADP detection FP assay kit (Bellbrook Labs, Madison, Wis.) using themanufacturer's recommended conditions for a 50 μM ATP reaction. Thefinal conditions for the assay were 50 mM HEPES, pH 7.2, 10 mM MgCl₂,7.5 mM tripotassium citrate, 2 mM DTT, 0.1 mg/mL BSA, 30 μM acetyl-CoA,50 μM ATP, and 10 mM KHCO₃. Typically, a 10 μl reaction was run for 120min at 25° C., and 10 μl of Transcreener stop and detect buffer wasadded and the combination incubated at room temp for an additional 1hour. The data was acquired on a Envision Fluorescence reader(Perkinelmer) using a 620 excitation Cy5 FP general dual mirror, 620excitation Cy5 FP filter, 688 emission (S) and a 688 (P) emissionfilter.

[3] Preparation of rhACC2. Human ACC2 inhibition was measured usingpurified recombinant human ACC2 (hrACC2). Briefly, a full length Cytomaxclone of ACC2 was purchased from Cambridge Bioscience Limited and wassequenced and subcloned into PCDNA5 FRT TO-TOPO (Invitrogen, Carlsbad,Calif.). The ACC2 was expressed in CHO cells by tetracycline inductionand harvested in 5 liters of DMEM/F12 with glutamine, biotin, hygromycinand blasticidin with 1 μg/mL tetracycline (Invitrogen, Carlsbad,Calif.). The conditioned medium containing ACC2 was then applied to aSoftlink Soft Release Avidin column (Promega, Madison, Wis.) and elutedwith 5 mM biotin. 4 mgs of ACC2 were eluted at a concentration of 0.05mg/mL (determined by A280) with an estimated purity of 95% (determinedby A280). The purified ACC2 was dialyzed in 50 mM Tris, 200 mM NaCl, 4mM DTT, 2 mM EDTA, and 5% glycerol. The pooled protein was frozen andstored at −80° C., with no loss of activity upon thawing. Formeasurement of ACC2 activity and assessment of ACC2 inhibition, testcompounds were dissolved in DMSO and added to the rhACC2 enzyme as a 5×stock with a final DMSO concentration of 1%.

[4] Measurement of human ACC2 inhibition. hACC2 was assayed in a Costar#3676 (Costar, Cambridge, Mass.) 384-well plate using the TranscreenerADP detection FP assay kit (Bellbrook Labs, Madison, Wis.) using themanufacturer's recommended conditions for a 50 uM ATP reaction. Thefinal conditions for the assay were 50 mM HEPES, pH 7.2, 5 mM MgCl₂, 5mM tripotassium citrate, 2 mM DTT, 0.1 mg/mL BSA, 30 μM acetyl-CoA, 50μM ATP, and 8 mM KHCO₃. Typically, a 10 μl reaction was run for 50 minat 25° C., and 10 μl of Transcreener stop and detect buffer was addedand the combination incubated at room temp for an additional 1 hour. Thedata was acquired on an Envision Fluorescence reader (Perkinelmer) usinga 620 excitation Cy5 FP general dual mirror, 620 excitation Cy5 FPfilter, 688 emission (S) and a 688 (P) emission filter.

The results using the recombinant hACC1 and recombinant hACC2Transcreener assays described above are summarized in the table belowfor the Compounds of Formula (I) exemplified in the Examples above. Allof the examples in both assays were run with a minimum of n=3.

hACC1 hACC2 Example (nM) (nM) 1 176 184 2 59.6 84.1 3 5400 2260 4 116 535 163 114 6 260 115 7 113 74.2 8 156 175 9 30.9 29.7 10 30.1 35.5 11 180268 12 152 94.5 13 221 143 14 139 76.1 15 6.11 7.88 16 9.75 11.6 17 15.621.6 18 4.5 14.6 19 837 487 20 355 300 21 627 507 22 1010 685 23 1370420 24 3140 557 25 600 202 26 20.4 7.49 27 2910 1140 28 6.70 5.35 2913.7 6.16 30 16.1 18.8 31 23.1 54.8 32 31.8 12.5 33 16.3 8.7 34 55.842.6 35 32.6 12.5 36 44.4 29.3 37 6.6 3.2 38 33.5 19.9 39 29.1 29.8 4010.2 6.1 41 8.6 15.2 42 39.9 30.4 43 98.4 133 44 34.4 35.9 45 4.9 10.046 11.6 15.7 47 32.5 25.7 48 59.5 28.0 49 54.7 24.7

Acute In Vivo Assessment of ACC Inhibition in Experimental Animals

The ACC inhibitory activity of the compounds of the present inventioncan be confirmed in vivo by evaluation of their ability to reducemalonyl-CoA levels in liver and muscle tissue from treated animals.

Measurement of malonyl-CoA production inhibition in experimentalanimals. In this method, male Sprague-Dawley Rats, maintained onstandard chow and water ad libitum (225-275 g), were randomized prior tothe study. Animals were either fed, or fasted for 18 hours prior to thebeginning of the experiment. Two hours into the light cycle the animalswere orally dosed with a volume of 5 mL/kg, (0.5% methyl cellulose;vehicle) or with the appropriate compound (prepared in vehicle). Fedvehicle controls were included to determine baseline tissue malonyl-CoAlevels while fasted animals were included to determine the effectfasting had on malonyl-CoA levels. One hour after compoundadministration the animals were asphyxiated with CO₂ and the tissueswere removed. Specifically, blood was collected by cardiac puncture andplaced into BD Microtainer tubes containing EDTA (BD Biosciences, NJ),mixed, and placed on ice. Plasma was used to determine drug exposure.Liver and quadriceps were removed, immediately freeze-clamped, wrappedin foil and stored in liquid nitrogen.

Tissues were pulverized under liquid N₂ to ensure uniformity insampling. Malonyl-CoA was extracted from the tissue (150-200 mg) with 5volumes 10% tricarboxylic acid in Lysing Matrix A (MP Biomedicals, PN6910) in a FastPrep FP120 (Thermo Scientific, speed=5.5; for 45seconds). The supernatant containing malonyl-CoA was removed from thecell debris after centrifugation at 15000×g for 30 minutes (EppendorfCentrifuge 5402). Samples were stably frozen at −80 C until analysis iscompleted.

Analysis of malonyl CoA levels in liver and muscle tissue can beevaluated using the following methodology.

The method utilizes the following materials: Malonyl-CoA tetralithiumsalt and malonyl-¹³C₃-CoA trilithium salt which were purchased fromIsotec (Miamisburg, Ohio, USA), sodium perchlorate (Sigma, cat no.410241), trichloroacetic acid (ACROS, cat no. 42145), phosphoric acid(J.T. Baker, cat no. 0260-01), ammonium formate (Fluka, cat no. 17843),methanol (HPLC grade, J.T. Baker, cat no. 9093-33), and water (HPLCgrade, J.T. Baker, 4218-03) were used to make the necessary mobilephases. Strata-X on-line solid phase extraction columns, 25 μm, 20mm×2.0 mm I.D (cat no. 00M-S033-B0-CB) were obtained from Phenomenex(Torrance, Calif., USA). SunFire C18 reversed-phase columns, 3.5 μm, 100mm×3.0 mm I.D. (cat no. 186002543) were purchased from WatersCorporation (Milford, Mass., USA).

This method may be performed utilizing the following equipment.Two-dimensional chromatography using an Agilent 1100 binary pump, anAgilent 1100 quaternary pump and two Valco Chem inert 6-port twoposition valves. Samples were introduced via a LEAP HTC PAL auto samplerwith Peltier cooled stack maintained at 10° C. and a 204 sampling loop.The needle wash solutions for the autosampler are 10% trichloroaceticacid in water (w/v) for Wash 1 and 90:10 methanol:water for Wash 2. Theanalytical column (Sunfire) was maintained at 35° C. using a MicroTechScientific Micro-LC Column Oven. The eluent was analyzed on an ABI SciexAPI3000 triple quadrupole mass spectrometer with Turbo Ion Spray.

Two-dimensional chromatography was performed in parallel using distinctgradient elution conditions for on-line solid phase extraction andreversed-phase chromatography. The general design of the method was suchthat the first dimension was utilized for sample clean-up and capture ofthe analyte of interest followed by a brief coupling of both dimensionsfor elution from the first dimension onto the second dimension. Thedimensions were subsequently uncoupled allowing for gradient elution ofthe analyte from the second dimension for quantification whilesimultaneously preparing the first dimension for the next sample in thesequence. When both dimensions were briefly coupled together, the flowof the mobile phase in the first dimension was reversed for analyteelution on to the second dimension, allowing for optimal peak width,peak shape, and elution time.

The first dimension of the HPLC system utilized the Phenomenex strata-Xon-line solid phase extraction column and the mobile phase consisted of100 mM sodium perchlorate/0.1% (v/v) phosphoric acid for solvent A andmethanol for solvent B.

The second dimension of the HPLC system utilized the Waters SunFire C18reversed-phase column and the mobile phase consisted of 100 mM ammoniumformate for solvent A and methanol for solvent B. The initial conditionof the gradient was maintained for 2 minutes and during this time theanalyte was transferred to the analytical column. It was important thatthe initial condition was at a sufficient strength to elute the analytefrom the on-line SPE column while retaining it on the analytical.Afterwards, the gradient rose linearly to 74.5% A in 4.5 minutes beforea wash and re-equilibration step.

Mass spectrometry when coupled with HPLC can be a highly selective andsensitive method for quantitatively measuring analytes in complexmatrices but is still subject to interferences and suppression. Bycoupling a two dimensional HPLC to the mass spectrometer, theseinterferences were significantly reduced. Additionally, by utilizing theMultiple Reaction Monitoring (MRM) feature of the triple quadrupole massspectrometer, the signal-to-noise ratio was significantly improved.

For this assay, the mass spectrometer was operated in positive ion modewith a TurbolonSpray voltage of 2250V. The nebulizing gas was heated to450° C. The Declustering Potential (DP), Focusing Potential (FP), andCollision Energy (CE) were set to 60, 340, and 42 V, respectively.Quadrupole 1 (Q1) resolution was set to unit resolution with Quadrupole3 (Q3) set to low. The CAD gas was set to 8. The MRM transitionsmonitored were for malonyl CoA: 854.1→347.0 m/z (L. Gao et al. (2007) J.Chromatogr. B 853, 303-313); and for malonyl-¹³C₃-CoA: 857.1→350.0 m/zwith dwell times of 200 ms. The eluent was diverted to the massspectrometer near the expected elution time for the analyte, otherwiseit was diverted to waste to help preserve the source and improverobustness of the instrumentation. The resulting chromatograms wereintegrated using Analyst software (Applied Biosystems). Tissueconcentrations for malonyl CoA were calculated from a standard curveprepared in a 10% solution of trichloroacetic acid in water.

Samples comprising the standard curve for the quantification ofmalonyl-CoA in tissue extracts were prepared in 10% (w/v)trichloroacetic acid (TCA) and ranged from 0.01 to 1 pmol/μL.Malonyl-¹³C₃-CoA (final concentration of 0.4 pmol/μL) was added to eachstandard curve component and sample as an internal standard.

Six intra-assay quality controls were prepared; three from a pooledextract prepared from fasted animals and three from a pool made from fedanimals. These were run as independent samples spiked with 0, 0.1 or 0.3pmol/μL ¹²C-malonyl-CoA as well as malonyl-¹³C₃-CoA (0.4 pmol/μL). Eachintra-assay quality control contained 85% of aqueous tissue extract withthe remaining portion contributed by internal standard (0.4 pmol/μL) and¹²C-malonyl-CoA. Inter assay controls were included in each run; theyconsist of one fasted and one fed pooled sample of quadriceps and/or onefasted and one fed pooled sample of liver. All such controls are spikedwith malonyl-¹³C₃-CoA (0.4 pmol/μL).

What is claimed is:
 1. A method for treating a diabetes-related diseasein a human comprising the step of administering to the human in need ofsuch treatment a therapeutically effective amount of a compound ofFormula (I)

or a pharmaceutically acceptable salt thereof, wherein R¹ is(C₁-C₆)alkyl, (C₃-C₇)cycloalkyl, tetrahydrofuranyl or oxetanyl, whereinsaid (C₁-C₆)alkyl is optionally substituted with 1 to 3 substituentsindependently selected from (C₁-C₃)alkoxy, hydroxy, fluoro, phenyl,tetrahydrofuranyl or oxetanyl; R² is hydrogen, halo, (C₁-C₃)alkyl, orcyano; R³ are each independently hydrogen or (C₁-C₃)alkyl; L is a directbond or a (C₁-C₆)alkylene wherein one carbon of the (C₁-C₆)alkylene isoptionally replaced by —C(O)—, —C(O)NH—, —NHC(O)—, —O—, —S—, NH orN(C₁-C₃)alkyl; Z is CH₂ or O; A¹ and A² are each independently(C₆-C₁₀)aryl, 5 to 12 membered heteroaryl or 8 to 12 membered fusedheterocyclicaryl; wherein said (C₆-C₁₀)aryl, 5 to 12 membered heteroarylor 8 to 12 membered fused heterocyclicaryl are each optionallysubstituted with one to three substituents independently selected from(C₁-C₃)alkyl, (C₁-C₃)alkoxy, halo, amino, (C₁-C₃)alkylamino,di(C₁-C₃)alkylamino, hydroxy, cyano and amido wherein the alkyl portionof the (C₁-C₃)alkyl, (C₁-C₃)alkoxy, (C₁-C₃)alkylamino anddi(C₁-C₃)alkylamino are optionally substituted with one to five fluoro;and wherein one of A¹ or A² is substituted by CO₂R⁴, (C₁-C₆)CO₂R⁴,tetrazolyl or (C₁-C₆)tetrazolyl; and R⁴ is (C₁-C₈)alkyl,(C₃-C₈)cycloalkyl or (C₁-C₆)alkyl-(C₃-C₈)cycloalkyl.
 2. The method ofclaim 1, wherein the diabetes-related disease is selected fromnonalcoholic fatty liver disease, hepatic insulin resistance,hyperglycemia, metabolic syndrome, impaired glucose tolerance, diabeticneuropathy, diabetic nephropathy, diabetic retinopathy, obesity,dyslipidemia, hypertension, hyperinsulinemia, and insulin resistancesyndrome.
 3. The method of claim 1, wherein the diabetes-related diseaseis selected from nonalcoholic fatty liver disease, hepatic insulinresistance, hyperglycemia, and obesity.
 4. The method of claim 1,wherein the diabetes-related disease is nonalcoholic fatty liverdisease.
 5. The method of claim 1, further comprising the step ofadministering a second pharmaceutical agent, wherein the second agent isan anti-diabetic agent selected from the group consisting of metformin,acetohexamide, chlorpropamide, diabinese, glibenclamide, glipizide,glyburide, glimepiride, gliclazide, glipentide, gliquidone, glisolamide,tolazamide, tolbutamide, tendamistat, trestatin, acarbose, adiposine,camiglibose, emiglitate, miglitol, voglibose, pradimicin-Q, salbostatin,balaglitazone, ciglitazone, darglitazone, englitazone, isaglitazone,pioglitazone, rosiglitazone, troglitazone, exendin-3, exendin-4,trodusquemine, reservatrol, hyrtiosal extract, sitagliptin,vildagliptin, alogliptin and saxagliptin.
 6. The method of claim 1,wherein the compound of Formula (I) is where R¹ is isopropyl or t-butyl;and where R² is hydrogen; or a pharmaceutically acceptable salt thereof.7. The method of claim 1, wherein the compound of Formula (I) is whereA¹ is phenyl, pyridinyl, indazolyl, indolyl, benzoimidazolyl,pyrrolopyridinyl or pyrrolopyrimidinyl; each optionally substituted withone methyl, methoxy, methylamino or dimethylamino; or a pharmaceuticallyacceptable salt thereof.
 8. The method of claim 1, wherein the compoundof Formula (I) is where A¹ is phenyl, indolyl or benzoimidazolyloptionally substituted with methyl, or pyridinyl optionally substitutedwith methylamino or dimethylamino; or a pharmaceutically acceptable saltthereof.